Characteristic chromatogram method and identification of codonopsis pilosula (codonopsis pilosula) medicinal material, honey-bran fried codonopsis pilosula (codonopsis pilosula) decoction pieces and standard decoction and formula granules of honey-bran fried codonopsis pilosula (codonopsis pilosula) decoction pieces

Abstract

The invention provides honey and bran fried codonopsis pilosula (codonopsis pilosula) decoction pieces, standard decoction, formula granules and a UPLC characteristic spectrum construction method of a codonopsis pilosula (codonopsis pilosula) medicinal material, and the method comprises the following steps: adding water into the codonopsis pilosula (codonopsis pilosula) medicinal material, decocting, filtering, adding water into residues, and carrying out ultrasonic treatment to obtain a codonopsis pilosula (codonopsis pilosula) medicinal material solution to be detected; the method comprises the following steps: mixing honey and bran fried codonopsis pilosula (codonopsis pilosula) raw materials and a solvent, and performing ultrasonic treatment to obtain honey and bran fried codonopsis pilosula (codonopsis pilosula) liquid to be detected; determining the codonopsis pilosula (codonopsis pilosula) medicinal material liquid to be detected by adopting a high performance liquid chromatography to obtain the UPLC characteristic spectrum of the codonopsis pilosula (codonopsis pilosula) medicinal material; and determining the honey-bran-fried codonopsis pilosula (codonopsis pilosula) liquid to be detected by adopting a high performance liquid chromatography to obtain the UPLC characteristic spectrum of the honey-bran-fried codonopsis pilosula (codonopsis pilosula) raw material. According to the method, the acetonitrile-0. 2% formic acid is used as a mobile phase for gradient elution, the codonopsis pilosula glycoside I is used as a reference substance, the established characteristic chromatogram is good in repeatability and precision, and the method is stable and reliable.

Description

Technical Field

[0001] The present invention relates to the technical field of analysis and detection, and in particular to a method for constructing UPLC characteristic spectra of honey bran fried Codonopsis pilosula (Dangshen) decoction pieces, standard decoctions, formula granules and Codonopsis pilosula (Dangshen) medicinal materials. Background Art

[0002] Honey bran fried codonopsis (dangshen) is a processed product obtained by frying codonopsis (dangshen) with honey bran until yellow, and then sifting out the bran. Honey bran fried codonopsis (dangshen) standard decoction is a freeze-dried powder made from the processed medicinal materials according to a fixed preparation process; honey bran fried codonopsis (dangshen) formula granules are formula granules made from the processed medicinal materials and processed according to the main quality indicators of the standard decoction.

[0003] In order to ensure the uniformity and stability of the quality of medicinal materials, decoction pieces, standard decoctions and formula granules, the present invention establishes a new characteristic spectrum method to control their quality; and can clearly distinguish them from Codonopsis pilosula (Dangshen) and its preparations. Summary of the invention

[0004] In view of this, the technical problem to be solved by the present invention is to provide a method for constructing a UPLC characteristic spectrum of honey bran fried Codonopsis (Dongshen) slices, standard decoctions, formula granules and Codonopsis (Dongshen) medicinal materials. The UPLC characteristic spectrum method of honey bran fried Codonopsis (Dongshen) slices, standard decoctions and formula granules constructed by the present invention is stable and reliable, and the quality of honey bran fried Codonopsis (Dongshen) slices, standard decoctions and formula granules can be controlled. Codonopsis is described in the 2020 edition of the Chinese Pharmacopoeia as three base sources: Codonopsis, Codonopsis pilosula and Codonopsis pilosula. The objects studied in the present invention all use raw materials with Codonopsis as the base source. The Codonopsis expressed in brackets in the full text represents the base source of the object of study of the present invention.

[0005] The present invention establishes a UPLC characteristic spectrum method for Codonopsis pilosula (Dongshen) medicinal materials, honey bran fried Codonopsis pilosula (Dongshen) decoction pieces and their standard decoctions and formula granules, which provides a scientific identification basis for identifying Codonopsis pilosula (Dongshen) medicinal materials, honey bran fried Codonopsis pilosula (Dongshen) decoction pieces and their standard decoctions and formula granules, and can clearly distinguish them from Codonopsis pilosula and its preparations.

[0006] The present invention provides a method for constructing a UPLC characteristic spectrum of honey bran fried Codonopsis pilosula (Dangshen) decoction pieces, standard decoctions, formula granules and / or Codonopsis pilosula (Dangshen) medicinal materials, comprising:

[0007] A) boiling Codonopsis pilosula (Dangshen) medicinal material with water, filtering, and ultrasonically treating the residue with water to obtain a Codonopsis pilosula (Dangshen) medicinal material test solution;

[0008] The raw material of honey bran fried codonopsis (dangshen) is extracted by solvent to obtain a honey bran fried codonopsis (dangshen) test solution; the raw material of honey bran fried codonopsis (dangshen) includes honey bran fried codonopsis (dangshen) decoction pieces, standard decoction, and formula granules;

[0009] B) using high performance liquid chromatography to measure the Codonopsis pilosula (Dangshen) medicinal material solution to obtain a UPLC characteristic spectrum of the Codonopsis pilosula (Dangshen) medicinal material;

[0010] The honey bran fried codonopsis (dangshen) test solution is measured by high performance liquid chromatography to obtain a UPLC characteristic spectrum of the honey bran fried codonopsis (dangshen) raw material;

[0011] The chromatographic conditions of the high performance liquid chromatography method are as follows: the chromatographic column is a C18 column; the mobile phase A is acetonitrile, the mobile phase B is 0.2% formic acid, and the elution is gradient;

[0012] The gradient elution is specifically:

[0013] 0-15min, phase A: 0-2%, phase B: 100-98%;

[0014] 15-30min, phase A: 2%-15%, phase B: 98%-85%;

[0015] 30-40min, phase A: 15%-20%, phase B: 85%-80%;

[0016] 40-50min, phase A: 20%-30%, phase B: 80%-70%.

[0017] The method for constructing UPLC characteristic spectrum of honey bran fried Codonopsis pilosula (Dongshen) slices, standard decoction, formula granules and Codonopsis pilosula (Dongshen) medicinal materials provided by the present invention firstly boils the Codonopsis pilosula (Dongshen) medicinal materials with water, filters them, and ultrasonically treats the residue with water to obtain a Codonopsis pilosula (Dongshen) medicinal material test solution; the mass volume ratio of the Codonopsis pilosula (Dongshen) medicinal materials boiled with water is 2g:100mL; the time for boiling the Codonopsis pilosula (Dongshen) medicinal materials with water is 20 to 30min, preferably 30min; the ultrasonic power is 600w, the frequency is 40KHz, and the ultrasonic time is 20 to 30min.

[0018] The raw material of stir-fried codonopsis pilosula (Dongshen) with honey bran and a solvent is mixed and ultrasonically treated to obtain a test solution of stir-fried codonopsis pilosula (Dongshen); the raw material of stir-fried codonopsis pilosula (Dongshen) with honey bran includes honey bran stir-fried codonopsis pilosula (Dongshen) decoction pieces, standard decoctions, and formula granules;

[0019] When the raw material of honey bran fried codonopsis (dangshen) is honey bran fried codonopsis slices, the pretreatment is as follows: honey bran fried codonopsis (dangshen) slices are decocted with water, filtered, the residue is treated with ultrasound with water, cooled, shaken, filtered, and the decocted time is 20 to 30 minutes; preferably 30 minutes; the ultrasonic power is 600w, the frequency is 40KHz, and the ultrasonic time is 20 to 30 minutes;

[0020] When the raw material of honey bran fried Codonopsis (Codonopsis pilosula) is honey bran fried Codonopsis pilosula (Codonopsis pilosula) standard decoction or formula granules, the pretreatment is: add water to the honey bran fried Codonopsis pilosula (Codonopsis pilosula) standard decoction or formula granules for ultrasonic treatment, cool, shake well, and filter to obtain; the ultrasonic power is 600w, the frequency is 40KHz, and the ultrasonic time is 20 to 30min; preferably 30min; the mass volume ratio of honey bran fried Codonopsis pilosula (Codonopsis pilosula) standard decoction or formula granules and water is 2g:25mL.

[0021] The present invention adopts the above extraction solvent, the chromatographic peak information volume is large and the effect is good.

[0022] It also includes the preparation of reference substances and reference medicinal material solutions:

[0023] Take the Codonopsis pilosula I reference substance and dissolve it in methanol to obtain a reference substance solution;

[0024] Take Codonopsis pilosula (Dangshen) reference medicinal material, add water to boil, filter, add water to the residue for ultrasonic treatment, cool, shake well, filter, take the filtrate, and obtain the reference medicinal material reference solution;

[0025] The reference substance and reference medicinal material reference substance solutions are determined by high performance liquid chromatography to obtain chromatograms of the reference substance and the reference medicinal material reference substance, respectively; and based on the chromatograms of the reference substance and the reference medicinal material reference substance, the components of honey bran fried Codonopsis pilosula (Codonopsis pilosula) decoction pieces, standard decoction, formula granules and Codonopsis pilosula medicinal material UPLC characteristic spectra are qualitatively determined.

[0026] The mobile phase A of the present invention is acetonitrile, the mobile phase B is 0.2% formic acid solution, and the elution is performed by gradient.

[0027] The gradient elution of the present invention is preferably specifically:

[0028] 0-15min, phase A: 0-2%, phase B: 100-98%;

[0029] 15-30min, phase A: 2%-15%, phase B: 98%-85%;

[0030] 30-40min, phase A: 15%-20%, phase B: 85%-80%;

[0031] 40-50min, phase A: 20%-30%, phase B: 80%-70%.

[0032] The present invention has good baseline separation under the above elution gradient, good separation of each peak and stable baseline.

[0033] C18 column, specifications: 1.6μm~1.9μm, 2.1×150mm; column temperature: 30℃.

[0034] The chromatographic column of the present invention is preferably: HypersilGOLD TM a Q 2.1×150 mm, 1.9 μm; ACQUITY BEHC182.1×150mm,1.6μm; Shim-pack GIST-HP C18-AQ2.1×150mm,1.7μm.

[0035] The chromatographic column of the present invention has symmetrical chromatographic peaks, good separation and moderate peak elution time under the above 30°C condition.

[0036] The mobile phase flow rate is preferably 0.2 ml / min.

[0037] The present invention found that the chromatographic peaks were better separated, the peak shape was more symmetrical, and the separation degree was moderate at a flow rate of 0.2 ml / min, which was the most preferred solution.

[0038] The detection wavelength of the present invention is preferably 268 nm.

[0039] The inventors found that at 268 nm, the chromatographic information is rich, the chromatographic peak information is large, each component has good absorption, the response value is moderate, the peak separation is good, and the baseline is stable.

[0040] The injection volume of the present invention is preferably 1 μL.

[0041] The beneficial effects of the present invention are that under one liquid chromatography condition, the substance groups of honey bran fried Codonopsis pilosula (Dongshen) decoction pieces, standard decoctions, formula granules and Codonopsis pilosula (Dongshen) medicinal materials are controlled by fingerprints, and the fingerprints are located by Codonoside I; the cost of detection can be greatly reduced and qualitative detection can be achieved.

[0042] The similarity of UPLC characteristic spectra of honey bran fried Codonopsis pilosula slices, standard decoction and its formula granules was evaluated by using a traditional Chinese medicine chromatographic fingerprint similarity evaluation system, and a UPLC standard characteristic spectrum of honey bran fried Codonopsis pilosula (Codonopsis pilosula) slices, standard decoction and its formula granules consisting of 9 characteristic peaks was obtained, wherein Peak 7 (S): Codonoside I; Peak 8: Codonoside glycoside nin; Peak 9: Codonoside glycoside; in the honey bran fried Codonopsis pilosula (Codonopsis pilosula) raw material characteristic spectrum, Codonoside I peak was taken as the reference peak S peak, and the relative retention time of each characteristic peak and the S peak was calculated, and the relative retention time was within ±10% of the specified value, and the specified values ​​were: 0.23 (peak 1), 0.26 (peak 2), 0.31 (peak 3), 0.35 (peak 4), 0.42 (peak 5), 0.81 (peak 6), 1.14 (peak 8), and 1.27 (peak 9), respectively.

[0043] The similarity of the UPLC characteristic spectrum of Codonopsis pilosula (Dongshen) medicinal material was evaluated by using the similarity evaluation system of traditional Chinese medicine chromatographic fingerprint, and a UPLC standard characteristic spectrum of Codonopsis pilosula (Dongshen) medicinal material consisting of 10 characteristic peaks was obtained, wherein Peak 7 (S): Codonoside I; Peak 8: Codonopsis linine; Peak 9: Codonopsis linine; in the characteristic spectrum of Codonopsis pilosula medicinal material, the Codonopsis linine I peak was used as the reference peak S peak, and the relative retention time of each characteristic peak and the S peak was calculated, and the relative retention time was within ±10% of the specified value, and the specified values ​​were: the specified values ​​were: 0.23 (peak 1), 0.26 (peak 2), 0.31 (peak 3), 0.35 (peak 4), 0.42 (peak 5), 0.63 (peak 6), 0.81 (peak 7), 1.14 (peak 9), and 1.27 (peak 10).

[0044] Quality judgment standard: Take honey bran fried Codonopsis (Codonopsis) slices, standard decoction, formula granules and Codonopsis (Codonopsis) medicinal material samples, and operate according to the same method as above to obtain the characteristic spectra of honey bran fried Codonopsis (Codonopsis) slices, standard decoction and its formula granules. The standard characteristic spectra and sample characteristic spectra of honey bran fried Codonopsis (Codonopsis) slices, standard decoction and its formula granules are analyzed using the "Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicine" 2012 edition of the National Pharmacopoeia Committee, and the similarity is greater than 0.90.

[0045] The present invention also provides a method for identifying honey-fried dangshen (codonopsis) pieces, standard decoctions, formula granules and dangshen (codonopsis) medicinal materials, characterized in that the identification is carried out by the method described in any one of claims 1 to 9; the UPLC characteristic spectrum of dangshen medicinal materials has 10 peaks; the UPLC characteristic spectrum of honey-fried dangshen (codonopsis) pieces and its standard decoctions and formula granules has 9 peaks; the dangshen (codonopsis) medicinal materials have 6 more peaks than the honey-fried dangshen raw materials, so the two can be identified.

[0046] The UPLC characteristic spectrum method of honey bran fried Codonopsis pilosula (Dongshen) decoction pieces, standard decoction and formula granules thereof of the present invention and the peaks identified from the chromatographic peaks are: Peak 7 (S): Codonoside I; Peak 8: Codonoside glycoside nin; Peak 9: Codonoside glycoside; The UPLC characteristic spectrum method of Codonopsis pilosula (Dongshen) medicinal material and the peaks identified from the chromatographic peaks are: Peak 8 (S): Codonoside I; Peak 9: Codonoside glycoside nin; Peak 10: Codonoside glycoside;

[0047] The UPLC characteristic spectrum of Codonopsis pilosula (Dangshen) medicinal material has 10 peaks; the UPLC characteristic spectrum of Codonopsis pilosula (Dangshen) decoction pieces stir-fried with honey bran and its standard decoction and formula granules has 9 peaks;

[0048] The medicinal material Codonopsis pilosula (Dangshen) has 10 peaks, which is 6 more than the honey bran fried Codonopsis pilosula (Dangshen) decoction pieces and its standard decoction and formula granules. It can clearly distinguish Codonopsis pilosula (Dangshen) and honey bran fried Codonopsis pilosula (Dangshen).

[0049] The method provided by the present invention can effectively monitor the quality of different batches of honey bran fried codonopsis (dangshen) decoction pieces, standard decoctions, formula granules and codonopsis medicinal materials, so that their quality is stable. The method has the characteristics of high precision and good reproducibility, which is conducive to comprehensive monitoring of product quality.

[0050] The honey bran stir-fried Codonopsis pilosula (Dongshen) decoction pieces, standard decoction, formula granules and Codonopsis pilosula (Dongshen) medicinal material characteristic maps established by the present invention take Codonopsis pilosula glycoside I as a reference, focus on the order of each characteristic peak and the correlation with the medicinal materials and intermediate products, can comprehensively evaluate the overall quality features of the products, and the method is scientific and reliable.

[0051] The newly-created characteristic spectrum method of the present invention is simple and easy to operate in the preparation method of the test sample, and relatively more characteristic peaks can be identified. Accurate and reliable characteristic spectrum detection can be performed on honey bran fried Codonopsis (Dongshen) pieces, standard decoctions, formula granules and Codonopsis (Dongshen) medicinal materials. The authenticity and quality consistency and stability of honey bran fried Codonopsis (Dongshen) pieces, standard decoctions, formula granules and Codonopsis (Dongshen) medicinal materials can be effectively detected and controlled. Provide a basis for effectively controlling and more comprehensively evaluating the quality of honey bran fried Codonopsis (Dongshen) pieces, standard decoctions, formula granules and Codonopsis (Dongshen) medicinal materials. Ensure the uniformity and stability of the quality of honey bran fried Codonopsis (Dongshen) pieces, standard decoctions, formula granules and Codonopsis (Dongshen) medicinal materials.

[0052] The invention provides a method for constructing UPLC characteristic spectrum of honey bran fried Codonopsis (Codonopsis) slices, standard decoctions, formula granules and Codonopsis (Codonopsis) medicinal materials, comprising: A) boiling Codonopsis medicinal materials with water, filtering, adding water to the residue for ultrasonic treatment to obtain a Codonopsis (Codonopsis) medicinal material test solution; mixing the honey bran fried Codonopsis (Codonopsis) raw materials and a solvent for ultrasonic treatment to obtain a honey bran fried Codonopsis (Codonopsis) test solution; the honey bran fried Codonopsis (Codonopsis) raw materials include honey bran fried Codonopsis (Codonopsis) slices, standard decoctions and formula granules; B) determining the Codonopsis (Codonopsis) medicinal material test solution by high performance liquid chromatography to obtain a UPLC characteristic spectrum of Codonopsis (Codonopsis) medicinal materials; The test solution is determined by high performance liquid chromatography to obtain the UPLC characteristic spectrum of the raw material of codonopsis pilosula (codonopsis pilosula) fried with honey bran; the chromatographic conditions of the high performance liquid chromatography are: the chromatographic column is a C18 column; the mobile phase A is acetonitrile, the mobile phase B is 0.2% formic acid, and the gradient elution is performed; the gradient elution is specifically as follows: 0-15min, phase A: 0-2%, phase B: 100-98%; 15-30min, phase A: 2%-15%, phase B: 98%-85%; 30-40min, phase A: 15%-20%, phase B: 85%-80%; 40-50min, phase A: 20%-30%, phase B: 80%-70%. The invention adopts high performance liquid chromatography, selects acetonitrile-0.2% formic acid as mobile phase for gradient elution, takes codonopsis pilosula I as reference, establishes UPLC characteristic spectra of honey bran fried codonopsis pilosula (dangshen) decoction pieces, standard decoction, formula granules and codonopsis pilosula (dangshen) medicinal materials, has good repeatability and precision, and the method is stable and reliable, and can control and identify the quality of honey bran fried codonopsis pilosula (dangshen) decoction pieces, standard decoction and formula granules. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] Figure 1 Codonopsis pilosula (Dangshen) medicinal material characteristics atlas;

[0054] Figure 2 Comparative characteristic spectrum of Codonopsis pilosula (Dangshen);

[0055] Figure 3 Investigation on the sample preparation method of Codonopsis pilosula (Dangshen) slices fried with honey bran;

[0056] Figure 4 Investigation of solvents for extracting Codonopsis pilosula (Dangshen) slices fried with honey bran;

[0057] Figure 5 Investigation on the amount of solvent added to honey bran fried Codonopsis pilosula (Dangshen) slices;

[0058] Figure 6 Investigation on the extraction time of honey bran fried Codonopsis pilosula (Dangshen) slices;

[0059] Figure 7 Chromatographic peak identification of characteristic spectrum of Codonopsis pilosula (Dangshen) slices stir-fried with honey bran;

[0060] Figure 8 Investigation of different chromatographic columns for honey bran fried Codonopsis pilosula (Dangshen) slices;

[0061] Fig. 9 Surveys with different instruments;

[0062] Fig.10 Characteristic spectrum of honey bran fried Codonopsis pilosula (Dangshen) decoction pieces;

[0063] Fig.11 Comparative characteristic spectrum of honey bran fried Codonopsis pilosula (Dangshen) decoction pieces;

[0064] Fig.12 It is the characteristic atlas of Codonopsis pilosula (Dangshen);

[0065] Fig.13 This is a comparative characteristic spectrum of Codonopsis pilosula (Dangshen);

[0066] Fig.14 Investigation on the sample preparation method of standard decoction of Codonopsis pilosula (Dangshen) stir-fried with honey bran;

[0067] Fig.15 Investigation of solvents for extracting standard decoction of Codonopsis pilosula (Dangshen) stir-fried with honey bran;

[0068] Fig.16 Investigation on the amount of solvent added to the standard decoction of Codonopsis pilosula (Dangshen) stir-fried with honey bran;

[0069] Fig.17 Extraction time investigation;

[0070] Fig.18 Chromatographic peak identification of standard decoction of Codonopsis pilosula (Dangshen) stir-fried with honey bran;

[0071] Fig.19 Investigation of different chromatographic columns for the standard decoction of Codonopsis pilosula (Dangshen) stir-fried with honey bran;

[0072] Fig. 20 Investigation of different instruments for the standard decoction of Codonopsis pilosula (Dangshen) stir-fried with honey bran;

[0073] Fig.21 Characteristic spectrum of standard decoction of Codonopsis pilosula (Dangshen) (Dangshen) stir-fried with honey bran;

[0074] Fig. 22 Comparative characteristic spectrum of standard decoction of Codonopsis pilosula (Dangshen) stir-fried with honey bran;

[0075] Fig.23 Chromatograms of different wavelengths of honey bran stir-fried Codonopsis pilosula (Dangshen) formula granules;

[0076] Fig.24 Mobile phase selection;

[0077] Fig.25Investigation of column temperature of honey bran stir-fried Codonopsis pilosula (Dangshen) formula granules;

[0078] Fig.26 Investigation of the particle flow rate of honey bran stir-fried Codonopsis pilosula (Dangshen) formula;

[0079] Fig. 27 Investigation on the delayed release of honey bran stir-fried Codonopsis pilosula (Dangshen) formula granules;

[0080] Fig.28 Investigation of sample preparation methods;

[0081] Fig.29 Investigation of extraction solvents;

[0082] Fig.30 Investigation of solvent addition amount;

[0083] Fig.31 Extraction time investigation;

[0084] Fig.32 Chromatographic peak identification of honey bran stir-fried Codonopsis pilosula (Dangshen) formula granules;

[0085] Fig.33 Investigation of different chromatographic columns for the formula granules of honey bran fried codonopsis (Dangshen);

[0086] Fig.34 Surveys with different instruments;

[0087] Fig.35 Characteristic spectrum of honey bran stir-fried Codonopsis pilosula (Dangshen) formula granules (MFCDS-KL02);

[0088] Fig.36 Characteristic spectrum of honey bran stir-fried Codonopsis pilosula (Dangshen) formula granules (MFCDS-KL03);

[0089] Fig.37 Characteristic spectrum of honey bran stir-fried Codonopsis pilosula (Dangshen) formula granules (MFCDS-KL03);

[0090] Fig.38 Comparative characteristic spectrum of honey bran stir-fried Codonopsis pilosula (Dangshen) formula granules;

[0091] Fig.39 It is the PLS-DA score graph of each batch of Codonopsis pilosula (Dangshen) medicinal materials and honey bran fried Codonopsis pilosula (Dangshen) decoction pieces;

[0092] Fig.40 The control fingerprint of the honey bran fried codonopsis (Dangshen) formula granules of comparative example 1;

[0093] Fig.41 Characteristic spectra of the medicinal materials, decoction pieces, standard soup and granules of the honey bran fried codonopsis (dangshen) formula granules of comparative example 1;

[0094] Fig.42The invention discloses an overlay of characteristic spectra of the honey bran stir-fried Codonopsis pilosula (Dangshen) medicinal materials, decoction pieces, standard soup and granules. DETAILED DESCRIPTION

[0095] The present invention provides a method for constructing UPLC characteristic spectra of honey bran fried Codonopsis pilosula (Dongshen) slices, standard decoctions, formula granules and Codonopsis pilosula (Dongshen) medicinal materials. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It is particularly important to point out that all similar replacements and modifications are obvious to those skilled in the art, and they all belong to the scope of protection of the present invention. The method and application of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the method and application of this article without departing from the content, spirit and scope of the present invention to realize and apply the technology of the present invention.

[0096] To further illustrate the present invention, the following is a detailed description of a UPLC characteristic spectrum construction method of a honey bran fried Codonopsis pilosula (Dongshen) decoction piece, a standard decoction, a formula granule and Codonopsis pilosula (Dongshen) medicinal material provided by the present invention in combination with the examples.

[0097] High performance liquid chromatograph: Agilent 1290 (instrument 1); Thermo Fisher Vanquish Flex (instrument 2); Shimadzu LC-40BX3 (instrument 3);

[0098] Electronic balance: ME204E / 02, MS205DU, XP26 (Mettler-Toledo Instrument Co., Ltd.);

[0099] Ultrapure water machine: Cell type 1810A (Shanghai Moller Scientific Instrument Co., Ltd.);

[0100] Ultrasonic cleaner: KQ600DB (600W, 40KHz; Kunshan Ultrasonic Instrument Co., Ltd.);

[0101] Chromatographic column: Hypersil GOLD TM a Q 2.1×150mm, 1.9μm (chromatographic column 1); ACQUITY BEHC182.1×150mm,1.6μm (chromatographic column 2); Shim-pack GIST-HP C18-AQ2.1×150mm,1.7μm (chromatographic column 3);

[0102] Acetonitrile and formic acid were of chromatographic grade, water was ultrapure water, and the other reagents were of analytical grade;

[0103] Codonopsis pilosula (China Food and Drug Inspection Institute, batch number: 111732-201908);

[0104] Codonopsis pilosula glycoside (Chengdu Pusi Biotechnology Co., Ltd., batch number: PS012906, purity: 98.98%);

[0105] Codonopsis pilosula I (Shandong Bokang Fine Chemical Co., Ltd., batch number: TC4172-221020);

[0106] Codonopsis pilosula (Dangshen) control drug (Batch No. 121057-202107, China Food and Drug Inspection Institute);

[0107] Codonopsis pilosula (Dangshen) medicinal materials (Batch numbers: DS01, DS02, DS03, DS04, DS05, DS06, DS07, DS08, DS09, DS10, DS11, DS12, DS13, DS-14, DS15, DS16, DS17, DS18)

[0108] Example 1 UPLC characteristic spectrum of Codonopsis pilosula (Dangshen) medicinal material

[0109] 1.1 Chromatographic conditions and system suitability test Octadecylsilane bonded silica gel was used as the filler (column length 150 mm, inner diameter 2.1 mm, particle size 1.6 μm); acetonitrile was used as mobile phase A, 0.2% formic acid solution was used as mobile phase B, and gradient elution was performed according to the provisions in the following table; the flow rate was 0.2 ml per minute; the column temperature was 30°C; the detection wavelength was 268 nm. The theoretical plate number calculated based on the codonopsis pilosula I peak should not be less than 5000.

[0110]

[0111]

[0112] Preparation of reference solution Take 2g of Codonopsis pilosula (Dangshen) reference medicinal material, place in a stoppered conical flask, add 100ml of water, boil for 30 minutes, filter, evaporate the filtrate, add 25ml of water to the residue, ultrasonically treat (power 600W, frequency 40kHz) for 30 minutes, cool, shake well, filter, and take the filtrate as the reference medicinal material reference solution. Take an appropriate amount of Codonopsis pilosula I reference substance, accurately weigh it, add methanol to make a solution containing 30μg per 1ml, as the reference substance reference solution.

[0113] Preparation of test solution Take 2 g of Codonopsis pilosula (Dangshen) powder (passed through No. 3 sieve), place in a stoppered conical flask, add 100 ml of water, boil for 30 minutes, filter, evaporate the filtrate to dryness, add 25 ml of water to the residue, ultrasonically treat (power 600 W, frequency 40 kHz) for 30 minutes, cool, shake well, filter, and take the filtrate to obtain the solution.

[0114] The determination method is to accurately pipette 1 μl of the reference solution and the test solution respectively, inject them into the liquid chromatograph, and determine the result.

[0115] 1.2 Determination of characteristic peaks and establishment of reference spectrum

[0116] This method was used to analyze the characteristic spectra of 18 batches of Codonopsis pilosula (Dangshen) medicinal material samples and calculate the relative retention time. Figure 1 , Table 1. Figure 1 Characteristic spectrum of medicinal material Codonopsis pilosula (Dangshen); Peak 8 (S): Codonoside I; Peak 9: Codonopsis pilosula glycoside; Peak 10: Codonopsis pilosula glycoside; (S1-S18 from bottom to top are: DS01, DS02, DS03, DS04, DS05, DS06, DS07, DS08, DS09, DS10, DS11, DS12, DS13, DS-14, DS15, DS16, DS17, DS18).

[0117] Table 1 Relative retention time of characteristic spectra of medicinal materials

[0118]

[0119] According to the principle that the relative retention time is stable and each batch of samples can be detected and the peak is relatively high, a total of 10 peaks with good repeatability were selected as characteristic peaks. The results showed that the relative retention time RSD of the 10 characteristic peaks of 18 batches of Codonopsis pilosula (Dangshen) medicinal materials was less than 10%.

[0120] Final regulations: There should be 10 characteristic peaks in the chromatogram of the test sample, and the retention time should correspond to the 10 characteristic peaks in the chromatogram of the reference medicinal material. Peak 8 should correspond to the retention time of the reference peak, and the peak corresponding to the peak of the reference peak of Codonopsis pilosula I is the S peak. The relative retention time of each characteristic peak and the S peak is calculated, and the relative retention time should be within the range of ±10% of the specified value. The specified values ​​are: 0.23 (peak 1), 0.26 (peak 2), 0.31 (peak 3), 0.35 (peak 4), 0.42 (peak 5), 0.63 (peak 6), 0.81 (peak 7), 1.14 (peak 9), 1.27 (peak 10).

[0121] The Chinese medicine chromatographic fingerprint similarity evaluation system (2012 version) was used to synthesize 18 batches of Codonopsis pilosula (Dangshen) medicinal materials, and a comparison map of the characteristic map of Codonopsis pilosula (Dangshen) medicinal materials was established. Figure 2 . Figure 2 This is the reference characteristic spectrum of Codonopsis pilosula (Dangshen); Peak 8 (S): Codonoside I; Peak 9: Codonopsis pilosula glycoside nin; Peak 10: Codonopsis pilosula glycoside.

[0122] In summary, the feature map method is:

[0123] Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler (column length 150 mm, inner diameter 2.1 mm, particle size 1.6 μm); acetonitrile was used as mobile phase A, 0.2% formic acid solution was used as mobile phase B, and gradient elution was performed according to the provisions in the following table; the flow rate was 0.2 ml per minute; the column temperature was 30°C; the detection wavelength was 268 nm. The theoretical plate number calculated based on the codonopsis pilosula I peak should not be less than 5000.

[0124]

[0125] Preparation of reference solution Take 2g of Codonopsis pilosula (Dangshen) reference medicinal material, place in a stoppered conical flask, add 100ml of water, boil for 30 minutes, filter, evaporate the filtrate, add 25ml of water to the residue, ultrasonically treat (power 600W, frequency 40kHz) for 30 minutes, cool, shake well, filter, and take the filtrate as the reference medicinal material reference solution. Take an appropriate amount of Codonopsis pilosula I reference substance, accurately weigh it, add methanol to make a solution containing 30μg per 1ml, as the reference substance reference solution.

[0126] Preparation of test solution Take 2 g of Codonopsis pilosula (Dangshen) powder (passed through No. 3 sieve), place in a stoppered conical flask, add 100 ml of water, boil for 30 minutes, filter, evaporate the filtrate to dryness, add 25 ml of water to the residue, ultrasonically treat (power 600 W, frequency 40 kHz) for 30 minutes, cool, shake well, filter, and take the filtrate to obtain the solution.

[0127] The determination method is to accurately pipette 1 μl of the reference solution and the test solution respectively, inject them into the liquid chromatograph, and determine the result.

[0128] There should be 10 characteristic peaks in the chromatogram of the test sample, and the retention times of the 10 characteristic peaks in the chromatogram of the reference medicinal material should correspond. Peak 8 should correspond to the retention time of the reference peak, and the peak corresponding to the Codonopsis I reference peak is the S peak. The relative retention time of each characteristic peak and the S peak is calculated, and the relative retention time should be within ±10% of the specified value. The specified values ​​are: 0.23 (peak 1), 0.26 (peak 2), 0.31 (peak 3), 0.35 (peak 4), 0.42 (peak 5), 0.63 (peak 6), 0.81 (peak 7), 1.14 (peak 9), 1.27 (peak 10). See Figure Figure 2 .

[0129] Example 2 UPLC characteristic spectrum of fried codonopsis pilosula (codonopsis pilosula) slices with honey bran Drafting instructions

[0130] 2.2.2.1 Instruments and materials

[0131] High performance liquid chromatograph: Agilent 1290 (instrument 1); Thermo Fisher Vanquish Flex (instrument 2); Shimadzu LC-40BX3 (instrument 3);

[0132] Electronic balance: ME204E / 02, MS205DU, XP26 (Mettler-Toledo Instrument Co., Ltd.);

[0133] Ultrapure water machine: Cell type 1810A (Shanghai Moller Scientific Instrument Co., Ltd.);

[0134] Ultrasonic cleaner: KQ600DB (600W, 40KHz; Kunshan Ultrasonic Instrument Co., Ltd.);

[0135] Chromatographic column Hypersil GOLD TM a Q 2.1×150mm, 1.9μm (chromatographic column 1); ACQUITY BEHC182.1×150mm,1.6μm (chromatographic column 2); Shim-pack GIST-HP C18-AQ2.1×150mm,1.7μm (chromatographic column 3);

[0136] Acetonitrile and formic acid were of chromatographic grade, water was ultrapure water, and the other reagents were of analytical grade;

[0137] Codonopsis pilosula (China Food and Drug Inspection Institute, batch number: 111732-201908);

[0138] Codonopsis pilosula glycoside (Chengdu Pusi Biotechnology Co., Ltd., batch number: PS012906, purity: 98.98%);

[0139] Codonopsis pilosula I (Shandong Bokang Fine Chemical Co., Ltd., batch number: TC4172-221020);

[0140] Codonopsis pilosula (Dangshen) control drug (Batch No. 121057-202107, China Food and Drug Inspection Institute);

[0141] Stir-fried Codonopsis pilosula (Dangshen) slices with honey bran (prepared by Sichuan New Green Pharmaceutical Technology Development Co., Ltd., batch numbers: MFCDS01, MFCDS02, MFCDS03, MFCDS04, MFCDS05, MFCDS06, MFCDS07, MFCDS08, MFCDS09, MFCDS10, MFCDS11, MFCDS12, MFCDS13, MFCDS-14, MFCDS15, MFCDS16, MFCDS17, MFCDS18)

[0142] Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler (column length 150 mm, inner diameter 2.1 mm, particle size 1.6 μm); acetonitrile was used as mobile phase A, 0.2% formic acid solution was used as mobile phase B, and gradient elution was performed according to the provisions in the following table; the flow rate was 0.2 ml per minute; the column temperature was 30°C; the detection wavelength was 268 nm. The theoretical plate number calculated based on the codonopsis pilosula I peak should not be less than 5000.

[0143]

[0144] Preparation of reference solution Take 2g of Codonopsis pilosula (Dangshen) reference medicinal material, place in a stoppered conical flask, add 100ml of water, boil for 30 minutes, filter, evaporate the filtrate, add 25ml of water to the residue, ultrasonically treat (power 600W, frequency 40kHz) for 30 minutes, cool, shake well, filter, and take the filtrate as the reference medicinal material reference solution. Take an appropriate amount of Codonopsis pilosula I reference substance, accurately weigh it, add methanol to make a solution containing 30μg per 1ml, as the reference substance reference solution.

[0145] Preparation of test solution: Take 2 g of the powder of the product (stir-fried Codonopsis pilosula (Dangshen) slices with honey bran) (passed through No. 3 sieve), put it in a stoppered conical flask, add 100 ml of water, boil for 30 minutes, filter, evaporate the filtrate to dryness, add 25 ml of water to the residue, ultrasonically treat (power 600 W, frequency 40 kHz) for 30 minutes, cool, shake well, filter, and take the filtrate to obtain the solution.

[0146] The determination method is to accurately pipette 1 μl of the reference solution and the test solution respectively, inject them into the liquid chromatograph, and determine the result.

[0147] 2.2.2.2 Chromatographic conditions and system suitability investigation

[0148] Characteristic chromatographic conditions were kept consistent with system suitability and characteristic profiling methods for particles and standard decoctions.

[0149] The chromatographic conditions and system suitability test of the characteristic spectrum of honey bran fried codonopsis (dangshen) slices were determined as follows: octadecylsilane bonded silica gel as filler (column length 150mm, inner diameter 2.1mm, particle size 1.6μm); acetonitrile as mobile phase A, 0.2% formic acid solution as mobile phase B, gradient elution as specified in the table below; flow rate 0.2ml per minute; column temperature 30℃; detection wavelength 268nm. The number of theoretical plates calculated based on the codonopsis glycoside I peak should not be less than 5000.

[0150]

[0151] 2.2.1.3 Preparation of test solution

[0152] 2.2.2.3.1 Investigation of sample preparation methods

[0153] Take 2g of honey bran fried Codonopsis pilosula (Dangshen) decoction powder (passed through No. 3 sieve) (batch number: MFCDS18), put it in a stoppered conical flask, add 25ml of methanol, seal it, and examine the extraction methods of the test sample when they are refluxed and ultrasonic, respectively. The extraction time is 30 minutes, let it cool, shake it well, filter it, and take the filtrate to obtain it; take another 2g, put it in a stoppered conical flask, add 100ml of water, boil it for 30 minutes, filter it, evaporate the filtrate, add 25ml of water to the residue, ultrasonically treat it (power 600W, frequency 40kHz) for 30 minutes, shake it well, filter it, and take the filtrate to obtain it. Figure 3 . Figure 3 Investigation on the sample preparation method of Codonopsis pilosula (Dangshen) slices stir-fried with honey bran; the results showed that the chromatographic peaks of ultrasonic extraction after boiling in water were more and consistent with the peaks of standard decoction, so the extraction method of the test sample was determined to be ultrasonic extraction after boiling in water.

[0154] 2.2.2.3.2 Investigation of extraction solvent

[0155] Take 2g of the powder (passed through No. 3 sieve) (batch number: MFCDS18), put it in a stoppered conical flask, add 100ml of water, boil for 30 minutes, filter, evaporate the filtrate, add 25ml of methanol, 30% methanol, 50% methanol, 80% methanol, and water to the residue, ultrasonically treat (power 600W, frequency 40kHz) for 30 minutes, shake well, filter, and take the filtrate to obtain. Figure 4 . Figure 4 Investigation on the extraction solvent of Codonopsis pilosula (Dangshen) slices stir-fried with honey bran; the results showed that the extraction efficiency was higher when the extraction solvent was water, and the extraction solvent was temporarily set as water.

[0156] 2.2.2.3.3 Investigation of solvent addition amount

[0157] Take 2g of the powder (passed through No. 3 sieve) (batch number: MFCDS18), put it in a stoppered conical flask, add 100ml of water, boil for 30 minutes, filter, evaporate the filtrate, add 10ml, 25ml, and 50ml of water to the residue, respectively, treat with ultrasound (power 600W, frequency 40kHz) for 30 minutes, shake well, filter, and take the filtrate to obtain. Figure 5 . Figure 5 The amount of solvent added to the honey bran fried Codonopsis pilosula (Dangshen) slices was investigated; the results showed that when the amount of extraction solvent added was 25 ml, the peak shape and separation of each chromatographic peak were better, and the peak size was appropriate, so 25 ml of solvent was selected.

[0158] 2.2.2.3.4 Extraction time investigation

[0159] Take 2g of the powder (passed through No. 3 sieve) (batch number: MFCDS18), put it in a stoppered conical flask, add 100ml of water, boil for 30 minutes, filter, evaporate the filtrate, add 25ml of water to the residue, ultrasonically treat (power 600W, frequency 40kHz) for 15 minutes, 30 minutes, and 60 minutes respectively, shake well, filter, and take the filtrate to obtain. Figure 6 . Figure 6 The extraction time of honey bran fried Codonopsis pilosula (Dangshen) slices was investigated. The results showed that the extraction was complete when the extraction time was 30 minutes. Therefore, the extraction time was determined to be 30 minutes.

[0160] In summary, the preparation method of the test solution of honey bran fried Codonopsis pilosula (Dangshen) slices was determined as follows: take 2 g of the powder of the product (passed through a No. 3 sieve), put it in a stoppered conical flask, add 100 ml of water, boil for 30 minutes, filter, evaporate the filtrate to dryness, add 25 ml of water to the residue, ultrasonically treat (power 600 W, frequency 40 kHz) for 30 minutes, shake well, filter, and take the filtrate.

[0161] 2.2.2.4 Methodological investigation

[0162] 2.2.2.4.1 Chromatographic peak identification

[0163] Preparation of test solution: According to the experimental conditions proposed above, prepare the test solution of honey bran fried Codonopsis pilosula (Dangshen) slices.

[0164] Preparation of reference solution: Take 2g of Codonopsis pilosula (Dangshen) reference medicinal material, place in a stoppered conical flask, add 100ml of water, boil for 30 minutes, filter, evaporate the filtrate, add 25ml of water to the residue, ultrasonically treat (power 600W, frequency 40kHz) for 30 minutes, shake well, filter, filter, and take the filtrate as the reference medicinal material reference solution. Take appropriate amounts of Codonopsis pilosula I, Codonopsis pilosulae glycosides, and Codonopsis pilosulae glycosides, accurately weigh, add 80% methanol to make a solution containing 30μg per 1ml, as the reference solution.

[0165] Preparation of negative control solution: According to the experimental conditions proposed above, a negative control solution of fried Codonopsis pilosula (Dangshen) slices without honey and bran was prepared.

[0166] Position the characteristic spectrum peaks of honey bran fried Codonopsis pilosula (Dangshen) slices. Figure 7 . Figure 7 Characteristic spectrum of honey bran stir-fried Codonopsis (Dangshen) slices chromatographic peak identification; Peak 7 (S) is Codonoside I, Peak 8 is Codonopsis arginine, Peak 9 is Codonopsis arginine. In the following methodological investigation, 9 characteristic peaks in the sample were investigated. Codonopsis (Dangshen) control medicinal material has one more characteristic peak than honey bran stir-fried Codonopsis (Dangshen) slices, indicating the change of chemical composition during the processing process.

[0167] 2.2.2.4.2 Precision test

[0168] Take the test solution of honey bran fried codonopsis (Dangshen) slices (batch number: MFCDS18), inject 6 times continuously according to the proposed experimental method, 1μl each time, and calculate the retention time of each characteristic peak. See Table 2.

[0169] Table 2 Precision investigation - retention time

[0170]

[0171] The results showed that the retention time RSD of each characteristic peak of the sample ranged from 0.02% to 0.19%, indicating that the instrument had good precision.

[0172] Precisely weigh 6 portions of honey bran stir-fried Codonopsis pilosula (Dangshen) slices (batch number: MFCDS18), prepare and measure according to the proposed experimental method, and calculate the relative retention time of each characteristic peak. See Table 3.

[0173] Table 3 Repeatability study - relative retention time

[0174]

[0175] The results showed that the RSD range of the relative retention time of the six samples was 0.00% to 1.59%, indicating that the method had good repeatability.

[0176] 2.2.2.4.4 Inspection by different personnel and at different times

[0177] Based on the experimental conditions proposed above, different personnel (A, B) accurately weighed 1 portion of honey bran fried Codonopsis pilosula (Dangshen) slices (batch number: MFCDS18) at different times (T1, T2) to prepare the test sample for determination. See Table 4.

[0178] Table 4 Personnel and time inspection-relative retention time

[0179]

[0180] The results showed that when different personnel measured the same sample at different times, the RSD of the relative retention time of each characteristic peak was 0.00% to 1.95%, and the method had good stability.

[0181] 2.2.2.4.5 Study on the durability of chromatographic columns

[0182] Based on the experimental conditions proposed above, the following were used: Hypersil GOLD TM a Q2.1×150mm,1.9μm (chromatographic column 1); ACQUITY BEHC18 2.1×150mm,1.6μm (chromatographic column 2); Shim-packGIST-HP C18-AQ2.1×150mm,1.7μm (chromatographic column 3)3 were used for analysis and investigation. The results are shown in Figure 8 , Table 5. Figure 8 Investigation of different chromatographic columns for honey bran fried Codonopsis pilosula (Dangshen) slices;

[0183] Table 5 Column durability investigation - relative retention time

[0184]

[0185]

[0186] The results showed that the RSD of the relative retention time of the characteristic peaks was between 1.41% and 30.59% when the three chromatographic columns were used to detect the samples. The durability of the chromatographic columns in this method was poor, and the separation degree of chromatographic column 1 was better. Therefore, chromatographic column 1 was recommended.

[0187] 2.2.2.4.6 Instrument durability inspection

[0188] Based on the experimental conditions proposed above, honey bran fried codonopsis (dangshen) slices (batch number: MFCDS18) were weighed to prepare the test solution, and the test was performed on instrument 1, instrument 2, and instrument 3 respectively. The results are shown in Fig. 9 , Table 6. Fig. 9 Different instrument surveys

[0189] Table 6 Instrument durability investigation - relative retention time

[0190]

[0191] The results showed that when the three instruments were used to detect the test samples, the RSD of the relative retention time of each characteristic peak was 0.51% to 8.22%, indicating that the instrument durability of this method was good.

[0192] 2.2.2.4.7 Stability study

[0193] Based on the above experimental conditions, take the same test solution and measure it at 0h, 2h, 6h, 12h, 18h and 24h respectively. See Table 7.

[0194] Table 7 Stability Study-Retention Time

[0195]

[0196] The results showed that the RSD of the characteristic peak retention time was between 0.03% and 1.34%, and the sample solution was stable within 24 hours.

[0197] In summary, the RSD of the relative retention time of each characteristic peak meets the requirements in the above investigations, and the method is good. The above 9 characteristic peaks were included in the subsequent investigation.

[0198] 2.2.2.4.8 Determination of characteristic peaks and establishment of reference spectrum

[0199] This method was used to analyze the characteristic spectra of 18 batches of honey bran fried Codonopsis pilosula (Dangshen) slices, and the relative retention time and relative peak area ratio were calculated. Fig.10 , Table 8. Fig.10 Characteristic spectrum of honey bran fried Codonopsis pilosula (Dangshen) slices; Peak 7 (S): Codonoside I; Peak 8: Codonoside glycoside nin; Peak 9: Codonoside glycoside (from bottom to top S1-S18 are: MFCDS01, MFCDS02, MFCDS03, MFCDS04, MFCDS05, MFCDS06, MFCDS07, MFCDS08, MFCDS09, MFCDS10, MFCDS11, MFCDS12, MFCDS13, MFCDS-14, MFCDS15, MFCDS16, MFCDS17, MFCDS18).

[0200] Table 8 Relative retention time of characteristic spectra of medicinal materials

[0201]

[0202]

[0203] According to the principle of stable relative retention time and the fact that each batch of samples can be detected and the peak is relatively high, a total of 9 peaks with good durability were selected as characteristic peaks. According to the results of the methodological investigation and the verification results of 18 batches of decoction pieces, the theoretical plate number calculated based on the codonopsis pilosula I peak should be no less than 5000.

[0204] Final regulations: 9 characteristic peaks should be present in the chromatogram of the test sample, and the retention time should correspond to the 9 characteristic peaks in the chromatogram of the reference medicinal material. Peak 7 should correspond to the retention time of the reference peak, and the peak corresponding to the peak of the reference peak of Codonopsis pilosula I is the S peak. The relative retention time of each characteristic peak and the S peak is calculated, and the relative retention time should be within the range of ±10% of the specified value. The specified values ​​are: 0.23 (peak 1), 0.26 (peak 2), 0.31 (peak 3), 0.35 (peak 4), 0.42 (peak 5), 0.81 (peak 6), 1.14 (peak 8), 1.27 (peak 9).

[0205] The Chinese medicine chromatographic fingerprint similarity evaluation system (2012 version) was used to synthesize 18 batches of honey bran fried Codonopsis pilosula (Dangshen) slices, and a comparison spectrum of the characteristic spectrum of honey bran fried Codonopsis pilosula (Dangshen) slices was established. Fig.11 . Fig.11Comparative characteristic spectrum of honey bran fried Codonopsis pilosula (Dangshen) slices; Peak 7 (S): Codonoside I; Peak 8: Codonoside glycoside nin; Peak 9: Codonoside glycoside

[0206] In summary, the feature map method is:

[0207] Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler (column length 150 mm, inner diameter 2.1 mm, particle size 1.6 μm); acetonitrile was used as mobile phase A, 0.2% formic acid solution was used as mobile phase B, and gradient elution was performed according to the provisions in the following table; the flow rate was 0.2 ml per minute; the column temperature was 30°C; the detection wavelength was 268 nm. The theoretical plate number calculated based on the codonopsis pilosula I peak should not be less than 5000.

[0208]

[0209] Preparation of reference solution Take 2g of Codonopsis pilosula (Dangshen) reference medicinal material, place in a stoppered conical flask, add 100ml of water, boil for 30 minutes, filter, evaporate the filtrate, add 25ml of water to the residue, ultrasonically treat (power 600W, frequency 40kHz) for 30 minutes, cool, shake well, filter, and take the filtrate as the reference medicinal material reference solution. Take an appropriate amount of Codonopsis pilosula I reference substance, accurately weigh it, and add methanol to make a solution containing 30μg per 1ml.

[0210] Preparation of test solution: Take 2 g of the product powder (passed through No. 3 sieve), place in a stoppered conical flask, add 100 ml of water, boil for 30 minutes, filter, evaporate the filtrate to dryness, add 25 ml of water to the residue, ultrasonically treat (power 600 W, frequency 40 kHz) for 30 minutes, shake well, filter, and take the filtrate to obtain the product.

[0211] The determination method is to accurately pipette 1 μl of the reference solution and the test solution respectively, inject them into the liquid chromatograph, and determine the result.

[0212] There should be 9 characteristic peaks in the chromatogram of the test sample, and the retention time should correspond to the 9 characteristic peaks in the chromatogram of the reference medicinal material. Peak 7 should correspond to the retention time of the reference peak, and the peak corresponding to the Codonopsis I reference peak is the S peak. The relative retention time of each characteristic peak and the S peak is calculated, and the relative retention time should be within ±10% of the specified value. The specified values ​​are: 0.23 (peak 1), 0.26 (peak 2), 0.31 (peak 3), 0.35 (peak 4), 0.42 (peak 5), 0.81 (peak 6), 1.14 (peak 8), 1.27 (peak 9). See Figure Fig.11 .

[0213] 18 batches of Codonopsis pilosula (Dangshen) medicinal materials feature maps Fig.12 , Fig.12It is the characteristic spectrum of the medicinal material Codonopsis pilosula (Dangshen); Peak 8 (S): Codonoside I; Peak 9: Codonopsis pilosula glycoside nin; Peak 10: Codonopsis pilosula glycoside; S1-S18 are: DS01, DS02, DS03, DS04, DS05, DS06, DS07, DS08, DS09, DS10, DS11, DS12, DS13, DS-14, DS15, DS16, DS17, DS18.

[0214] 18 batches of Codonopsis pilosula (Dangshen) medicinal materials synthesis comparison chart Fig.13 , Fig.13 This is the reference characteristic spectrum of Codonopsis pilosula (Dangshen); Peak 8 (S): Codonoside I; Peak 9: Codonopsis pilosula glycoside nin; Peak 10: Codonopsis pilosula glycoside.

[0215] Example 3 UPLC characteristic spectrum of standard decoction of Codonopsis pilosula (Dangshen) fried with honey bran

[0216] 2.2.3.1 Instruments and materials

[0217] High performance liquid chromatograph: Agilent 1290 (instrument 1); Thermo Fisher Vanquish Flex (instrument 2); Shimadzu LC-40BX3 (instrument 3);

[0218] Electronic balance: ME204E / 02, MS205DU, XP26 (Mettler-Toledo Instrument Co., Ltd.);

[0219] Ultrapure water machine: Cell type 1810A (Shanghai Moller Scientific Instrument Co., Ltd.);

[0220] Ultrasonic cleaner: KQ600DB (600W, 40KHz; Kunshan Ultrasonic Instrument Co., Ltd.);

[0221] Chromatographic column: Hypersil GOLD TM a Q2.1×150mm, 1.9μm (chromatographic column 1); ACQUITY BEH C18 2.1×150mm, 1.6μm (chromatographic column 2); Shim-pack GIST-HP C18-AQ 2.1×150mm, 1.7μm (chromatographic column 3);

[0222] Acetonitrile and formic acid were of chromatographic grade, water was ultrapure water, and the other reagents were of analytical grade;

[0223] Codonopsis pilosula (China Food and Drug Inspection Institute, batch number: 111732-201908);

[0224] Codonopsis pilosula glycoside (Chengdu Pusi Biotechnology Co., Ltd., batch number: PS012906, purity: 98.98%);

[0225] Codonopsis pilosula I (Shandong Bokang Fine Chemical Co., Ltd., batch number: TC4172-221020);

[0226] Codonopsis pilosula (Dangshen) control drug (Batch No. 121057-202107, China Food and Drug Inspection Institute);

[0227] Standard decoction of Codonopsis pilosula (Dangshen) stir-fried with honey bran (prepared by Sichuan New Green Pharmaceutical Technology Development Co., Ltd., batch numbers: MFCDS-BT301, MFCDS-BT401, MFCDS-BT402, MFCDS-BT403, MFCDS-BT404, MFCDS-BT405, MFCDS-BT406, MFCDS-BT407, MFCDS-BT408, MFCDS-BT409, MFCDS-BT410, MFCDS-BT411, MFCDS-BT412, MFCDS-BT413, MFCDS-BT414, MFCDS-BT415, MFCDS-BT416, MFCDS-BT417).

[0228] Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler (column length 150 mm, inner diameter 2.1 mm, particle size 1.6 μm); acetonitrile was used as mobile phase A, 0.2% formic acid solution was used as mobile phase B, and gradient elution was performed according to the provisions in the following table; the flow rate was 0.2 ml per minute; the column temperature was 30°C; the detection wavelength was 268 nm. The theoretical plate number calculated based on the codonopsis pilosula I peak should not be less than 5000.

[0229]

[0230] Preparation of reference solution Take 2g of Codonopsis pilosula (Dangshen) reference medicinal material, place in a stoppered conical flask, add 100ml of water, boil for 30 minutes, filter, evaporate the filtrate, add 25ml of water to the residue, ultrasonically treat (power 600W, frequency 40kHz) for 30 minutes, cool, shake well, filter, and take the filtrate as the reference medicinal material reference solution. Take an appropriate amount of Codonopsis pilosula I reference substance, accurately weigh it, and add methanol to make a solution containing 30μg per 1ml.

[0231] Preparation of test solution: Take 2 g of the product (Standard decoction of Codonopsis pilosula (Dangshen) fried with honey bran), place it in a stoppered conical flask, add 25 ml of water, ultrasonically treat (power 600 W, frequency 40 kHz) for 30 minutes, cool, shake well, filter, and take the filtrate to obtain the solution.

[0232] The determination method is to accurately pipette 1 μl of the reference solution and the test solution respectively, inject them into the liquid chromatograph, and determine the result.

[0233] 2.2.3.2 Chromatographic conditions and system suitability test

[0234] The material composition basis of the standard decoction and the finished granules is consistent, and the characteristic chromatographic conditions are consistent with the system suitability and the characteristic chromatographic method of the granules.

[0235] Chromatographic conditions and system suitability test: octadecylsilane bonded silica gel is used as filler (column length 150 mm, inner diameter 2.1 mm, particle size 1.6 μm); acetonitrile is used as mobile phase A, 0.2% formic acid solution is used as mobile phase B, and gradient elution is performed according to the provisions in the following table; flow rate is 0.2 ml per minute; column temperature is 30°C; detection wavelength is 268 nm. The number of theoretical plates calculated based on the codonopsis pilosula I peak should not be less than 5000.

[0236]

[0237] 2.2.3.3 Investigation of sample preparation methods

[0238] 2.2.3.3.1 Investigation of extraction methods

[0239] Take 2g of this product (Standard Decoction of Codonopsis pilosula (Dangshen) Stir-fried with Honey Bran) (Batch No.: MFCDS-BT414), put it in a stoppered conical flask, add 25ml of methanol, seal it tightly, and examine the extraction method of the test sample when it is refluxed and ultrasonic respectively, the extraction time is 30 minutes, cool, shake well, filter, and take the filtrate to obtain. Fig.14 . Fig.14 The sample preparation method of standard decoction of Codonopsis pilosula (Dangshen) stir-fried with honey bran was investigated. The results showed that there was little difference in the effect of reflux and ultrasonic extraction of the test sample, and the ultrasonic method was fast and simple, so the extraction method of the test sample was determined to be ultrasonic extraction.

[0240] 2.2.3.3.2 Investigation of extraction solvent

[0241] Take 2g of this product (Standard Decoction of Codonopsis pilosula (Dangshen) Stir-fried with Honey Bran) (Batch No.: MFCDS-BT414), place it in a stoppered conical flask, and test the extraction solvents of the test sample as methanol, 30% methanol, 50% methanol, 80% methanol, and water 25ml each, seal it tightly, and treat it with ultrasound (power 600W, frequency 40kHz) for 30 minutes, cool it, shake it well, filter it, and take the filtrate to obtain it. Fig.15 . Fig.15 The extraction solvent of the standard decoction of Codonopsis pilosula (Dangshen) stir-fried with honey bran was investigated; the results showed that the extraction efficiency was higher when the extraction solvent was water, and the extraction solvent was temporarily set as water.

[0242] 2.2.3.3.3 Investigation of solvent addition amount

[0243] Take 2g of standard decoction of Codonopsis pilosula (Dangshen) stir-fried with honey bran (batch number: MFCDS-BT414), place it in a stoppered conical flask, add 25ml, 50ml, and 100ml of water respectively for investigation, seal it tightly, and treat it with ultrasound (power 600W, frequency 40kHz) for 30 minutes, let it cool, shake it well, filter it, and take the filtrate to obtain it. Fig.16 ; Fig.16 The amount of solvent added to the standard decoction of fried Codonopsis pilosula (Dangshen) with honey bran was investigated; the results showed that when the amount of extraction solvent added was 25 ml, the peak shape and separation of each chromatographic peak were better, and the peak size was appropriate, so 25 ml of solvent was selected.

[0244] 2.2.3.3.4 Extraction time investigation

[0245] Take 2g of standard decoction of Codonopsis pilosula (Dangshen) stir-fried with honey bran (batch number: MFCDS-BT414), put it in a stoppered conical flask, add 25ml of water, stopper it, and ultrasonically treat it (power 600W, frequency 40kHz) for 15 minutes, 30 minutes, and 60 minutes respectively, cool it, shake it well, filter it, and take the filtrate to obtain it. Fig.17 . Fig.17 The results of the extraction time showed that the extraction was complete when the extraction time was 30 minutes. Therefore, the extraction time was determined to be 30 minutes.

[0246] In summary, the preparation method of the test solution of the standard decoction of Codonopsis pilosula (Dangshen) fried with honey bran was determined as follows: take 2 g of the product, put it in a stoppered conical flask, add 25 ml of water, ultrasonically treat (power 600 W, frequency 40 kHz) for 30 minutes, cool, shake well, filter, and take the filtrate.

[0247] 2.2.3.4 Methodological investigation

[0248] 2.2.3.4.1 Chromatographic peak identification

[0249] Preparation of test solution: According to the experimental conditions proposed above, prepare the test solution of standard decoction of Codonopsis pilosula (Dangshen) fried with honey bran.

[0250] Preparation of reference solution: Take 2g of Codonopsis pilosula (Dangshen) as a reference medicinal material, place it in a stoppered conical flask, add 100ml of water, boil for 30 minutes, filter, evaporate the filtrate, add 25ml of water to the residue, ultrasonically treat (power 600W, frequency 40kHz) for 30 minutes, shake well, filter, and take the filtrate as a reference medicinal material reference solution. Separately take appropriate amounts of Codonopsis pilosula I, Codonopsis pilosulae glycosides, and Codonopsis pilosulae glycosides, accurately weigh them, and add methanol to prepare solutions containing 30μg per 1ml.

[0251] Preparation of negative control solution: According to the experimental conditions proposed above, a negative control solution of the standard decoction of fried Codonopsis pilosula (Dangshen) with bran lacking honey was prepared.

[0252] Positioning of characteristic peaks of the standard decoction of Codonopsis pilosula (Dangshen) with honey bran. Fig.18 . Fig.18 Chromatographic peak identification of standard decoction of Codonopsis pilosula stir-fried with honey bran; the results showed that peak 7 (S) was codonoside I, peak 8 was codonopsis linine, and peak 9 was codonopsis linine. In the following methodological investigation, 9 characteristic peaks in the sample were investigated. The control herb of Codonopsis pilosula (Dangshen) had one more characteristic peak than the standard decoction of Codonopsis pilosula stir-fried with honey bran, indicating the change of chemical composition during the processing process.

[0253] 2.2.3.4.2 Precision test

[0254] Take the standard decoction of honey bran fried codonopsis (Dangshen) (batch number: MFCDS-BT414) for the test solution, inject it 6 times continuously according to the proposed experimental method, 1 μl each time, and calculate the retention time and peak area of ​​each characteristic peak. See Table 9.

[0255] Table 9 Precision investigation - retention time

[0256]

[0257] The results showed that the RSD of the retention time of each characteristic peak was 0.02% to 0.24%, indicating that the instrument had good precision.

[0258] 2.2.3.4.3 Repeatability study

[0259] Weigh 6 portions of standard decoction of fried codonopsis pilosula with honey bran (batch number: MFCDS-BT414) accurately, prepare and measure according to the proposed experimental method, and calculate the relative retention time and relative peak area of ​​each characteristic peak. See Table 10.

[0260] Table 10 Repeatability study - relative retention time

[0261]

[0262] The results showed that the RSD of the relative retention time of the six samples was 0.00% to 1.59%, indicating that the method had good repeatability.

[0263] 2.2.3.4.4 Investigation by different personnel and time

[0264] Based on the above proposed experimental conditions, different personnel (A, B) accurately weighed the standard decoction of fried codonopsis pilosula (Dangshen) with honey bran (batch number: MFCDS-BT-230414) at different times (T1, T2) to prepare the test sample, and the relative retention time and relative peak area of ​​each characteristic peak were calculated. See Table 11.

[0265] Table 11 Personnel and time inspection-relative retention time

[0266]

[0267] The results showed that when different personnel measured the same sample at different times, the RSD of the relative retention time of each characteristic peak was 0.00% to 1.95%, and the method had good stability.

[0268] 2.2.3.4.5 Column durability investigation

[0269] Based on the above experimental conditions, the chromatographic columns were Hypersil GOLD TM a Q2.1×150mm,1.9μm (chromatographic column 1); ACQUITY BEH C18 2.1×150mm,1.6μm (chromatographic column 2); Shim-pack GIST-HP C18-AQ2.1×150mm,1.7μm (chromatographic column 3)3 were analyzed and investigated. The results are shown in Fig.19 , Table 12. Fig.19 Investigation of different chromatographic columns for standard decoction of fried codonopsis pilosula with honey bran

[0270] Table 12 Column investigation - relative retention time

[0271]

[0272] The results show that the RSD of the relative retention time of the characteristic peaks is between 1.41% and 28.76% when the three chromatographic columns are used to detect the samples. The durability of the chromatographic columns in this method is poor, and the separation degree of chromatographic column 1 is better. It is recommended to use chromatographic column 1.

[0273] 2.2.2.4.6 Instrument durability inspection

[0274] Based on the experimental conditions proposed above, analysis and investigation were carried out using Instrument 1, Instrument 2, and Instrument 3 respectively. The results are shown in Fig. 20 , Table 13. Fig. 20 Investigation of different instruments for the standard decoction of fried codonopsis pilosula with honey bran

[0275] Table 13 Column durability investigation - relative retention time

[0276]

[0277] The results showed that when the three instruments were used to detect the test samples, the RSD of the relative retention time of each characteristic peak was 0.51% to 8.22%, indicating that the instrument durability of this method was good.

[0278] 2.2.3.4.7 Stability study

[0279] Based on the above proposed experimental conditions, take the same test solution and measure it at 0h, 2h, 6h, 12h, 18h and 24h respectively. See Table 14.

[0280] Table 14 Stability Study-Retention Time

[0281]

[0282] The results showed that the RSD of the characteristic peak retention time was between 0.02% and 1.44%, and the sample solution was stable within 24 hours.

[0283] In summary, the RSD of the relative retention time of each characteristic peak meets the requirements in the above investigations, and the method is good. The above 9 characteristic peaks were included in the subsequent investigation.

[0284] 2.2.3.4.8 Determination of characteristic peaks and establishment of reference spectrum

[0285] This method was used to analyze the characteristic spectra of 18 batches of samples and calculate the relative retention time and relative peak area. Fig.21 , Table 15. Fig.21 Characteristic spectrum of standard decoction of Codonopsis pilosula (Dangshen) stir-fried with honey bran;

[0286] Peak 7 (S): Codonopsis ginsenoside I; Peak 8: Codonopsis ginsenoside nin; Peak 9: Codonopsis ginsenoside; S1-S18 from bottom to top are: MFCDS-BT301, MFCDS-BT401, MFCDS-BT402, MFCDS-BT403, MFCDS-BT404, MFCDS-BT405, MFCDS-BT406, MFCDS-BT407, MFCDS-BT408, MFCDS-BT409, MFCDS-BT410, MFCDS-BT411, MFCDS-BT412, MFCDS-BT413, MFCDS-BT414, MFCDS-BT415, MFCDS-BT416, MFCDS-BT417)

[0287] Table 15 Relative retention time of standard decoction of fried codonopsis pilosula (Dangshen) with honey bran

[0288]

[0289] According to the principle of stable relative retention time and the fact that each batch of samples can be detected and the peak is relatively high, a total of 9 peaks with good durability were selected as characteristic peaks. According to the results of the methodological investigation and the verification results of 18 batches of standard soup, the theoretical plate number calculated based on the codonopsis pilosula I peak should be no less than 5000.

[0290] Final regulations: 9 characteristic peaks should be present in the chromatogram of the test sample, and the retention time should correspond to the 9 characteristic peaks in the chromatogram of the reference medicinal material. Peak 7 should correspond to the retention time of the reference peak, and the peak corresponding to the peak of the reference peak of Codonopsis pilosula I is the S peak. The relative retention time of each characteristic peak and the S peak is calculated, and the relative retention time should be within the range of ±10% of the specified value. The specified values ​​are: 0.23 (peak 1), 0.26 (peak 2), 0.31 (peak 3), 0.35 (peak 4), 0.42 (peak 5), 0.81 (peak 6), 1.14 (peak 8), 1.27 (peak 9).

[0291] The similarity evaluation system of Chinese medicine chromatographic fingerprints (2012 version) was used to synthesize 18 batches of standard decoctions of Codonopsis pilosula (Dangshen) fried with honey bran, and a comparison spectrum of the characteristic spectrum of standard decoctions of Codonopsis pilosula (Dangshen) fried with honey bran was established. Fig. 22 . Fig. 22 Comparative characteristic spectrum of standard decoction of fried Codonopsis pilosula with honey bran; Peak 7 (S): Codonoside I; Peak 8: Codonopsis pilosula glycoside nin; Peak 9: Codonopsis pilosula glycoside.

[0292] In summary, the feature map method is:

[0293] Octadecylsilane bonded silica gel is used as filler (column length 150 mm, inner diameter 2.1 mm, particle size 1.6 μm); acetonitrile is used as mobile phase A, 0.2% formic acid solution is used as mobile phase B, and gradient elution is performed according to the provisions in the following table; the flow rate is 0.2 ml per minute; the column temperature is 30°C; the detection wavelength is 268 nm. The theoretical plate number calculated based on the codonopsis pilosula I peak should not be less than 5000.

[0294]

[0295] Preparation of reference solution Take 2g of Codonopsis pilosula (Dangshen) reference medicinal material, place in a stoppered conical flask, add 100ml of water, boil for 30 minutes, filter, evaporate the filtrate, add 25ml of water to the residue, ultrasonically treat (power 600W, frequency 40kHz) for 30 minutes, cool, shake well, filter, and take the filtrate as the reference medicinal material reference solution. Take an appropriate amount of Codonopsis pilosula I reference substance, accurately weigh it, and add methanol to make a solution containing 30μg per 1ml.

[0296] Preparation of test solution: Take 2 g of freeze-dried powder of standard decoction of Codonopsis pilosula (Dangshen) fried with honey bran, place it in a stoppered conical flask, add 25 ml of water, ultrasonically treat (power 600 W, frequency 40 kHz) for 30 minutes, cool, shake well, filter, and take the filtrate.

[0297] The determination method is to accurately pipette 1 μl of the reference solution and the test solution respectively, inject them into the liquid chromatograph, and determine the result.

[0298] There should be 9 characteristic peaks in the chromatogram of the test sample, and the retention time should correspond to the 9 characteristic peaks in the chromatogram of the reference medicinal material. Peak 7 should correspond to the retention time of the reference peak, and the peak corresponding to the Codonopsis I reference peak is the S peak. The relative retention time of each characteristic peak and the S peak is calculated, and the relative retention time should be within ±10% of the specified value. The specified values ​​are: 0.23 (peak 1), 0.26 (peak 2), 0.31 (peak 3), 0.35 (peak 4), 0.42 (peak 5), 0.81 (peak 6), 1.14 (peak 8), 1.27 (peak 9). See Figure Fig. 22 .

[0299] Example 4 UPLC characteristic spectrum of honey bran fried codonopsis (codonopsis) formula granules

[0300] 2.2.4.1 Instruments and materials

[0301] High performance liquid chromatograph: Agilent 1290 (instrument 1); Thermo Fisher Vanquish Flex (instrument 2); Shimadzu LC-40B X3 (instrument 3);

[0302] Electronic balance: ME204E / 02, MS205DU, XP26 (Mettler-Toledo Instrument Co., Ltd.);

[0303] Ultrapure water machine: Cell type 1810A (Shanghai Moller Scientific Instrument Co., Ltd.);

[0304] Ultrasonic cleaner: KQ600DB (600W, 40KHz; Kunshan Ultrasonic Instrument Co., Ltd.);

[0305] Chromatographic column: Hypersil GOLD TM a Q2.1×150mm, 1.9μm (chromatographic column 1); ACQUITY BEH C18 2.1×150mm, 1.6μm (chromatographic column 2); Shim-pack GIST-HP C18-AQ 2.1×150mm, 1.7μm (chromatographic column 3);

[0306] Acetonitrile and formic acid were of chromatographic grade, water was ultrapure water, and the other reagents were of analytical grade;

[0307] Codonopsis pilosula (China Food and Drug Inspection Institute, batch number: 111732-201908);

[0308] Codonopsis pilosula glycoside (Chengdu Pusi Biotechnology Co., Ltd., batch number: PS012906, purity: 98.98%);

[0309] Codonopsis pilosula I (Shandong Bokang Fine Chemical Co., Ltd., batch number: TC4172-221020);

[0310] Codonopsis pilosula (Dangshen) control drug (Batch No. 121057-202107, China Food and Drug Inspection Institute);

[0311] Honey bran stir-fried Codonopsis pilosula (Dangshen) formula granules (Sichuan New Green Pharmaceutical Technology Development Co., Ltd., batch numbers MFCDS-KL01, MFCDS-KL02, MFCDS-KL03, MFCDS-KL04)

[0312] Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler (column length 150 mm, inner diameter 2.1 mm, particle size 1.6 μm); acetonitrile was used as mobile phase A, 0.2% formic acid solution was used as mobile phase B, and gradient elution was performed according to the provisions in the following table; the flow rate was 0.2 ml per minute; the column temperature was 30°C; the detection wavelength was 268 nm. The theoretical plate number calculated based on the codonopsis pilosula I peak should not be less than 5000.

[0313]

[0314] Preparation of reference solution Take 2g of Codonopsis pilosula (Dangshen) reference medicinal material, place it in a stoppered conical flask, add 100ml of water, boil for 30 minutes, filter, evaporate the filtrate, add 25ml of water to the residue, ultrasonically treat (power 600W, frequency 40kHz) for 30 minutes, shake well, filter, and take the filtrate as the reference medicinal material reference solution. Take an appropriate amount of Codonopsis pilosula I reference substance, accurately weigh it, and add methanol to make a solution containing 30μg per 1ml.

[0315] Preparation of test solution: Take an appropriate amount of the product, grind it into powder, take 2 g, put it in a stoppered conical flask, add 25 ml of water, ultrasonically treat (power 600 W, frequency 40 kHz) for 30 minutes, cool, shake well, filter, and take the filtrate to obtain the solution.

[0316] The determination method is to accurately pipette 1 μl of the reference solution and the test solution respectively, inject them into the liquid chromatograph, and determine the result.

[0317] 2.2.4.2 Wavelength, column temperature, flow rate, mobile phase selection and delay test

[0318] 2.2.4.2.1 Wavelength selection

[0319] Based on the experimental conditions proposed above, the diode array detector was used to perform full-band scanning of the test solution, and the chromatograms of the test solution at wavelengths of 210nm, 230nm, 250nm, 268nm, 290nm, and 310nm were extracted respectively. Fig.23 . Fig.23Chromatograms of honey bran fried codonopsis pilosula granules at different wavelengths; the results showed that when the detection wavelength was 268nm, the chromatographic peak had a larger amount of information and the chromatogram baseline was more stable, so the detection wavelength was determined to be 268nm.

[0320] 2.2.4.2.2 Mobile phase selection

[0321] Based on the experimental conditions proposed above, the separation effects of four different mobile phases were investigated, namely: acetonitrile-0.2% formic acid, acetonitrile-0.2% phosphoric acid, acetonitrile-water, see Fig.24 . Fig.24 Selection of mobile phase; The results showed that the chromatogram baseline was relatively stable and there were more chromatographic peaks under the gradient elution condition of acetonitrile-0.2% formic acid solution, so the gradient elution of acetonitrile-0.2% formic acid solution was used as the mobile phase for the characteristic spectrum determination method of honey bran fried codonopsis (codonopsis) formula granules.

[0322] 2.2.4.2.3 Column temperature investigation

[0323] Based on the experimental conditions proposed above, the column temperatures were investigated at 25°C, 30°C, and 35°C. Fig.25 . Fig.25 The column temperature of honey bran fried codonopsis (dangshen) formula granules was investigated; the results showed that when the column temperature was 30℃, the chromatogram peak shape was more symmetrical, the separation was better, and the peaks were more complete. Therefore, the column temperature was determined to be 30℃.

[0324] 2.2.4.2.4 Flow rate investigation

[0325] Based on the experimental conditions proposed above, flow rates of 0.15ml / min, 0.2ml / min, 0.25ml / min, and 0.3ml / min were investigated. Fig.26 . Fig.26 The flow rate of the honey bran fried codonopsis (dangshen) formula granules was investigated; the results showed that when the flow rate was 0.2ml / min, the chromatogram peak shape was better and the separation was moderate. Therefore, the flow rate was determined to be 0.2ml / min.

[0326] 2.2.4.2.5 Delay Investigation

[0327] Based on the experimental conditions proposed above, a delay test was conducted. The results are shown in Fig. 27 . Fig. 27 Investigation on the delayed nature of the formula granules of Codonopsis pilosula (Dangshen) stir-fried with honey bran; the results showed that the sample had basically no chromatographic peak after 50 minutes, so the sample detection time was set at 50 minutes.

[0328] In summary, the chromatographic conditions and system suitability test for the characteristic spectrum of honey bran fried codonopsis (dangshen) formula granules were determined as follows: octadecylsilane bonded silica gel as filler (column length 150 mm, inner diameter 2.1 mm, particle size 1.6 μm); acetonitrile as mobile phase A, 0.2% formic acid solution as mobile phase B, gradient elution as specified in the table below; flow rate 0.2 ml per minute; column temperature 30 ° C; detection wavelength 268 nm. The number of theoretical plates calculated based on the codonopsis glycoside I peak should not be less than 5000.

[0329]

[0330] 2.2.4.3 Preparation of test solution

[0331] 2.2.4.3.1 Investigation of extraction methods

[0332] Take an appropriate amount of this product (batch number: MFCDS-KL01), grind it into powder, take 2g, put it in a stoppered conical flask, add 25ml of methanol, plug it tightly, and examine the extraction method of the test product when it is refluxed and ultrasonic, respectively. The extraction time is 30 minutes, cool, shake well, filter, and take the filtrate to obtain Fig.28 . Fig.28 The sample preparation method was investigated; the results showed that there was little difference in the effects of reflux and ultrasonic extraction of the test sample, and the ultrasonic method was fast and simple, so the extraction method of the test sample was determined to be ultrasonic extraction.

[0333] 2.2.4.3.2 Investigation of extraction solvent

[0334] Take an appropriate amount of this product (batch number: MFCDS-KL01), grind it into powder, take 2g, put it in a stoppered conical flask, and test the extraction solvents of the test product with methanol, 30% methanol, 50% methanol, 80% methanol, and water 25ml each, seal it tightly, and treat it with ultrasound (power 600W, frequency 40kHz) for 30 minutes, cool it, shake it well, filter it, and take the filtrate to obtain it. Fig.29 . Fig.29 Investigation of extraction solvent; the results showed that the chromatographic peaks obtained by using water as the extraction solvent had a large amount of information and good separation, and the extraction solvent was temporarily determined to be water.

[0335] 2.2.4.3.3 Investigation of solvent addition amount

[0336] Take an appropriate amount of this product (batch number: MFCDS-KL01), grind it into powder, take 2g, put it in a stoppered conical flask, add 25ml, 50ml, and 100ml of water respectively, seal it tightly, and treat it with ultrasound (power 600W, frequency 40kHz) for 30 minutes, cool it, shake it well, filter it, and take the filtrate to obtain it. Fig.30 . Fig.30The amount of solvent added was investigated; the results showed that when the amount of extraction solvent was 25 ml, the size of the chromatographic peak was more appropriate, so 25 ml of solvent was selected.

[0337] 2.2.4.3.4 Extraction time investigation

[0338] Take an appropriate amount of this product (batch number: MFCDS-KL01), grind it into powder, take 2g, put it in a stoppered conical flask, add 25ml of water, stopper it, and treat it with ultrasound (power 600W, frequency 40kHz). The extraction time of the test product is 15 minutes, 30 minutes, and 60 minutes respectively. Cool it, shake it well, filter it, and take the filtrate to obtain it. Fig.31 . Fig.31 The results of the extraction time showed that the extraction was complete when the extraction time was 30 minutes. Therefore, the extraction time was determined to be 30 minutes.

[0339] In summary, the preparation method of the test solution of the honey bran fried codonopsis (dangshen) formula granules was determined as follows: take an appropriate amount of the product, grind it into powder, take 2 g, put it in a stoppered conical flask, add 25 ml of water, ultrasonically treat (power 600 W, frequency 40 kHz) for 30 minutes, cool, shake well, filter, and take the filtrate to obtain the product.

[0340] 2.2.4.4 Methodological investigation

[0341] 2.2.4.4.1 Chromatographic peak identification

[0342] Preparation of test solution: According to the experimental conditions proposed above, prepare the test solution of honey bran fried codonopsis (Dangshen) formula granules.

[0343] Preparation of reference solution: Take 2g of Codonopsis pilosula (Dangshen) as a reference medicinal material, place it in a stoppered conical flask, add 100ml of water, boil for 30 minutes, filter, evaporate the filtrate, add 25ml of water to the residue, ultrasonically treat (power 600W, frequency 40kHz) for 30 minutes, shake well, filter, and take the filtrate as a reference medicinal material reference solution. Separately take appropriate amounts of Codonopsis pilosula I, Codonopsis pilosulae glycosides, and Codonopsis pilosulae glycosides, accurately weigh them, and add methanol to prepare solutions containing 30μg per 1ml.

[0344] Preparation of negative control solution: According to the experimental conditions proposed above, a negative control solution of the formula granules of stir-fried Codonopsis pilosula (Dangshen) with honey-deficient bran was prepared.

[0345] Positioning of characteristic spectrum peaks of honey bran fried codonopsis (dangshen) formula granules. Fig.32 . Fig.32Chromatographic peak identification of honey bran stir-fried Codonopsis pilosula (Dangshen) formula granules; the results showed that peak 7 (S) was codonoside I, peak 8 was codonopsis linine, and peak 9 was codonopsis linine. In the following methodological investigation, 9 characteristic peaks in the sample were investigated. The control drug Codonopsis pilosula (Dangshen) had one more characteristic peak than the honey bran stir-fried Codonopsis pilosula (Dangshen) formula granules, indicating the change of chemical composition during the processing process.

[0346] 2.2.4.4.2 Precision test

[0347] Take the test solution of honey bran fried codonopsis pilosula formula granules (batch number: MFCDS-KL01), inject 6 times continuously according to the proposed experimental method, 1 μl each time, and calculate the retention time and peak area of ​​each characteristic peak. See Table 16.

[0348] Table 16 Precision investigation - retention time

[0349]

[0350] The results showed that the RSD of the retention time of each characteristic peak was 0.03% to 0.23%, indicating that the instrument had good precision.

[0351] 2.2.4.4.3 Repeatability study

[0352] Precisely weigh 6 portions of honey bran stir-fried Codonopsis pilosula (Dangshen) formula granules (batch number: MFCDS-KL01) and prepare and measure them according to the proposed experimental method. See Table 17.

[0353] Table 17 Repeatability Study - Relative Retention Time

[0354]

[0355] The results showed that the RSD of the relative retention time of the six samples was 0.00% to 1.59%, indicating that the method had good repeatability.

[0356] 2.2.4.4.4 Investigation by different personnel and time

[0357] Based on the experimental conditions proposed above, different personnel (A, B) accurately weighed 1 portion of honey bran fried codonopsis (Dangshen) formula granules (batch number: MFCDS-KL01) at different times (T1, T2) to prepare the test sample for determination. See Table 18.

[0358] Table 18 Personnel and time inspection-relative retention time

[0359]

[0360] The results showed that when different personnel measured the same sample at different times, the RSD of the relative retention time of each characteristic peak was 0.00% to 1.95%, and the method had good stability.

[0361] 2.2.4.4.5 Study on the durability of chromatographic columns

[0362] Based on the above experimental conditions, the chromatographic columns were Hypersil GOLD TM a Q2.1×150mm,1.9μm (chromatographic column 1); ACQUITY BEHC18 2.1×150mm,1.6μm (chromatographic column 2); Shim-pack GIST-HP C18-AQ2.1×150mm,1.7μm (chromatographic column 3) were used for analysis and investigation. The results are shown in Fig.33 , Table 19. Fig.33 Investigation of different chromatographic columns for the formula granules of stir-fried codonopsis pilosula (Dangshen) with honey bran.

[0363] Table 19 Column durability study - relative retention time

[0364]

[0365] The results showed that the RSD of the relative retention time of the characteristic peaks was between 1.41% and 30.59% when the samples were detected using the above three chromatographic columns. The durability of the chromatographic columns of this method was poor, and the separation degree of chromatographic column 1 was better; chromatographic column 1 was recommended.

[0366] 2.2.2.4.6 Instrument durability inspection

[0367] Based on the experimental conditions proposed above, the analysis was carried out using Agilent 1290 (instrument 1); Thermo Fisher VanquishFlex (instrument 2); and Shimadzu LC-40BX3 (instrument 3). The results are shown in Fig.34 , Table 20. Fig.34 Different instrument surveys.

[0368] Table 20 Column durability study - relative retention time

[0369]

[0370] The results showed that when the three instruments were used to detect the test samples, the RSD of the relative retention time of each characteristic peak was 0.51% to 8.22%, indicating that the instrument durability of this method was good.

[0371] 2.2.4.4.7 Stability study

[0372] Based on the above proposed experimental conditions, take the same test solution and measure it at 0h, 2h, 6h, 9h, 12h, 18h and 24h respectively. See Table 21.

[0373] Table 21 Stability Study-Retention Time

[0374]

[0375] The results showed that the RSD of the characteristic peak retention time was between 0.04% and 1.43%, and the sample solution was stable within 24 hours.

[0376] In summary, the RSD of the relative retention time of each characteristic peak meets the requirements in the above investigations, and the method is good. The above 9 characteristic peaks were included in the subsequent investigation.

[0377] 2.2.4.4.8 Determination of characteristic peaks and establishment of reference spectrum

[0378] Using this method, characteristic spectrum analysis was performed on three batches of samples to calculate relative retention time and relative peak area. Figure 35-37 , Table 22. Fig.35 Characteristic spectrum of honey bran fried codonopsis (dangshen) formula granules (MFCDS-KL01); Peak 7 (S): codonoside I; Peak 8: codonopsis arginine; Peak 9: codonopsis arginine;

[0379] Fig.36 Characteristic spectrum of honey bran fried codonopsis (dangshen) formula granules (MFCDS-KL02); Peak 7 (S): codonoside I; Peak 8: codonopsis glycoside nin; Peak 9: codonopsis glycoside

[0380] Fig.37 Characteristic spectrum of honey bran fried Codonopsis pilosula (Dangshen) formula granules (MFCDS-KL03); Peak 7 (S): Codonoside I; Peak 8: Codonoside glycoside nin; Peak 9: Codonoside glycoside.

[0381] Table 22 Relative retention time of honey bran fried codonopsis (dangshen) formula granules

[0382]

[0383]

[0384] According to the principle of stable relative retention time and the fact that each batch of samples can be detected and the peaks are relatively high, a total of 9 peaks were selected as characteristic peaks. According to the results of the methodological investigation and the verification results of 3 batches of particles, the theoretical plate number calculated based on the codonopsis pilosula I peak should be no less than 5000.

[0385] Final regulations: 9 characteristic peaks should be present in the chromatogram of the test sample, and the retention time should correspond to the 9 characteristic peaks in the chromatogram of the reference medicinal material. Peak 7 should correspond to the retention time of the reference peak, and the peak corresponding to the peak of the reference peak of Codonopsis pilosula I is the S peak. The relative retention time of each characteristic peak and the S peak is calculated, and the relative retention time should be within the range of ±10% of the specified value. The specified values ​​are: 0.23 (peak 1), 0.26 (peak 2), 0.31 (peak 3), 0.35 (peak 4), 0.42 (peak 5), 0.81 (peak 6), 1.14 (peak 8), 1.27 (peak 9).

[0386] Three batches of honey bran stir-fried Codonopsis pilosula (Dangshen) formula granules were synthesized using the Chinese medicine chromatographic fingerprint similarity evaluation system (2012 version), and a comparison spectrum of the characteristic spectrum of honey bran stir-fried Codonopsis pilosula (Dangshen) formula granules was established. Fig.38 . Fig.38 Comparative characteristic spectrum of honey bran fried Codonopsis pilosula (Dangshen) formula granules; Peak 7 (S): Codonoside I; Peak 8: Codonoside glycoside nin; Peak 9: Codonoside glycoside.

[0387] In summary, the feature map method is:

[0388] Octadecylsilane bonded silica gel is used as filler (column length 150 mm, inner diameter 2.1 mm, particle size 1.6 μm); acetonitrile is used as mobile phase A, 0.2% formic acid solution is used as mobile phase B, and gradient elution is performed according to the provisions in the following table; the flow rate is 0.2 ml per minute; the column temperature is 30°C; the detection wavelength is 268 nm. The theoretical plate number calculated based on the codonopsis pilosula I peak should not be less than 5000.

[0389]

[0390] Preparation of reference solution Take 2g of Codonopsis pilosula (Dangshen) reference medicinal material, place it in a stoppered conical flask, add 100ml of water, boil for 30 minutes, filter, evaporate the filtrate, add 25ml of water to the residue, ultrasonically treat (power 600W, frequency 40kHz) for 30 minutes, shake well, filter, and take the filtrate as the reference medicinal material reference solution. Take an appropriate amount of Codonopsis pilosula I reference substance, accurately weigh it, and add methanol to make a solution containing 30μg per 1ml.

[0391] Preparation of test solution: Take an appropriate amount of the product, grind it into powder, take 2 g, put it in a stoppered conical flask, add 25 ml of water, ultrasonically treat (power 600 W, frequency 40 kHz) for 30 minutes, cool, shake well, filter, and take the filtrate to obtain the solution.

[0392] The determination method is to accurately pipette 1 μl of the reference solution and the test solution respectively, inject them into the liquid chromatograph, and determine the result.

[0393] There should be 9 characteristic peaks in the chromatogram of the test sample, and the retention time should correspond to the 9 characteristic peaks in the chromatogram of the reference medicinal material. Peak 7 should correspond to the retention time of the reference peak, and the peak corresponding to the Codonopsis I reference peak is the S peak. The relative retention time of each characteristic peak and the S peak is calculated, and the relative retention time should be within ±10% of the specified value. The specified values ​​are: 0.23 (peak 1), 0.26 (peak 2), 0.31 (peak 3), 0.35 (peak 4), 0.42 (peak 5), 0.81 (peak 6), 1.14 (peak 8), 1.27 (peak 9). See Figure Fig.38 .

[0394] Example 5

[0395] The 18 batches of Codonopsis pilosula (Dangshen) medicinal materials and 18 batches of honey bran fried Codonopsis pilosula (Dangshen) decoction pieces samples in this project were analyzed by PCA using the data statistical software SIMCA, with the peak areas of the 9 common peaks as variables. The results are shown in Fig.39 . Fig.39 PLS-DA score chart of each batch of Codonopsis pilosula (Dangshen) medicinal materials and honey bran fried Codonopsis pilosula (Dangshen) decoction pieces. Note: DS1~DS18 represent 18 batches of Codonopsis pilosula (Dangshen) medicinal materials; MFCDS1~MFCDS18 represent 18 batches of honey bran fried Codonopsis pilosula (Dangshen) decoction pieces; t in the horizontal and vertical axes represents the principal component in PLS-DA.

[0396] The cumulative contribution rate of the two principal components t1 and t2 was 95.0%, indicating that the model had good prediction performance. The results showed that PCA could distinguish Codonopsis pilosula (Dangshen) medicinal materials and honey bran fried Codonopsis pilosula (Dangshen) decoction pieces, Codonopsis pilosula (Dangshen) medicinal materials (DS1~DS18) were mainly distributed on the right side of the score graph, and honey bran fried Codonopsis pilosula (Dangshen) decoction pieces (MFCDS1~MFCDS18) were mainly distributed on the left side of the score graph.

[0397] The ultra-high performance liquid chromatography method for identifying the characteristic spectra of Codonopsis pilosula (Dongshen) medicinal materials, honey bran fried Codonopsis pilosula (Dongshen) decoction pieces, their standard decoctions and their formula granules constructed by the present invention can not only intuitively and quickly identify Codonopsis pilosula (Dongshen) medicinal materials and honey bran fried Codonopsis pilosula (Dongshen) decoction pieces in terms of the number of chromatographic peaks, but can also distinguish them by PCA analysis using data statistical software SIMCA. At the same time, the method of this project can also reflect the transmission of the characteristic spectra of the whole process of honey bran fried Codonopsis pilosula (Dongshen) formula granules from medicinal materials, decoction pieces, standard decoction to granules.

[0398] Comparative Example 1

[0399] 1.1 Preparation of test solution Take an appropriate amount of honey bran fried codonopsis pilosula granules sample, grind it into powder, take about 2 g, accurately weigh it, put it in a stoppered conical flask, accurately add 25 mL of methanol, stopper it, weigh it, and treat it with ultrasound for 30 min (power 250 w, frequency 40 kHz). Cool it, weigh it again, make up the lost mass with methanol, shake it well, filter it, and take the filtrate to obtain the test solution.

[0400] Preparation of reference solution: Take appropriate amount of codonopsis ginsenoside I, codonopsis ginsenoside nin, codonopsis ginsenoside and codonopsis ginsenoside alcohol reference substances, weigh them accurately, add methanol to make mixed solutions containing 10 μg of codonopsis ginsenoside I, codonopsis ginsenoside nin, codonopsis ginsenoside and codonopsis ginsenoside alcohol per 1 mL, as reference substance solutions.

[0401] Chromatographic conditions: Waters ACQUITY UPLC HSS T3 as chromatographic column (2.1mm×100mm, 1.8μm); acetonitrile as mobile phase A, 0.1% formic acid aqueous solution as mobile phase B, elution according to the gradient in Table 1; column temperature 30℃, flow rate 0.3mL / min, detection wavelength 268nm, injection volume 3μL. The theoretical plate number calculated based on the dangshen glycoside peak should not be less than 10.000.

[0402] The gradient is:

[0403]

[0404] Fig.40 For comparative example 1, the fingerprint of the honey bran fried codonopsis (dangshen) formula granules is compared. Fig.40 It can be seen that, excluding the solvent peak, there are only two peaks in the chromatogram. The above spectrum has poor reproducibility and the peak difference cannot be well distinguished. 1.2

[0406] The method of comparative example 1 was used to detect the characteristic spectrum of the medicinal materials, decoction pieces, standard soup and granules of honey bran fried codonopsis (codonopsis) formula granules, and the results are shown in Fig.41 ; Fig.41 Characteristic spectra of the medicinal materials, decoction pieces, standard soup and granules of the honey bran fried codonopsis (dangshen) formula granules of comparative example 1;

[0407] Depend on Fig.40 The results show that the chromatographic method in Comparative Example 1 cannot reflect the transfer process of the chromatographic peaks of the whole process of the honey bran fried codonopsis formula granules from medicinal materials, decoction pieces, standard soup to granules, while the chromatographic method constructed in the present invention can show the transfer process of the chromatographic peaks of the whole process of the honey bran fried codonopsis formula granules from medicinal materials, decoction pieces, standard soup to granules. Fig.42 ; Fig.42 The invention discloses an overlay of characteristic spectra of the honey bran stir-fried Codonopsis pilosula (Dangshen) medicinal materials, decoction pieces, standard soup and granules.

[0408] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for constructing a UPLC characteristic spectrum of honey bran fried Codonopsis pilosula (Dangshen) decoction pieces, standard decoctions, formula granules and / or Codonopsis pilosula (Dangshen) medicinal materials, comprising: A) boiling Codonopsis pilosula (Dangshen) medicinal material with water, filtering, and ultrasonically treating the residue with water to obtain a Codonopsis pilosula (Dangshen) medicinal material test solution; The raw material of honey bran fried codonopsis (dangshen) is extracted by solvent to obtain a honey bran fried codonopsis (dangshen) test solution; the raw material of honey bran fried codonopsis (dangshen) includes honey bran fried codonopsis (dangshen) decoction pieces, standard decoction, and formula granules; B) using high performance liquid chromatography to measure the Codonopsis pilosula (Dangshen) medicinal material solution to obtain a UPLC characteristic spectrum of the Codonopsis pilosula (Dangshen) medicinal material; The honey bran fried codonopsis (dangshen) test solution is measured by high performance liquid chromatography to obtain a UPLC characteristic spectrum of the honey bran fried codonopsis (dangshen) raw material; The chromatographic conditions of the high performance liquid chromatography method are as follows: the chromatographic column is a C18 column; the mobile phase A is acetonitrile, the mobile phase B is 0.2% formic acid, and the elution is gradient; The gradient elution is specifically: 0-15min, phase A: 0-2%, phase B: 100-98%; 15-30min, phase A: 2%-15%, phase B: 98%-85%; 30-40min, phase A: 15%-20%, phase B: 85%-80%; 40-50min, phase A: 20%-30%, phase B: 80%-70%.

2. The method according to claim 1, characterized in that It also includes the preparation of reference substances and reference medicinal material solutions: Take the Codonopsis pilosula I reference substance and dissolve it in methanol to obtain a reference substance solution; Take Codonopsis pilosula reference medicinal material, add water to boil, filter, add water to the residue for ultrasonic treatment, cool, shake well, filter, take the filtrate, and obtain a reference medicinal material reference solution; The reference substance and reference medicinal material reference substance solutions are determined by high performance liquid chromatography to obtain chromatograms of the reference substance and the reference medicinal material reference substance, respectively; and based on the chromatograms of the reference substance and the reference medicinal material reference substance, the components of honey bran fried Codonopsis pilosula (Dongshen) decoction pieces, standard decoction, formula granules and UPLC characteristic spectra of Codonopsis pilosula (Dongshen) medicinal materials are qualitatively determined.

3. The method according to claim 1, characterized in that The chromatographic column is a C18 column with specifications of 1.6-1.9 μm and 2.1×150 mm; the column temperature is 30° C.

4. The method according to claim 1, characterized in that The flow rate of the mobile phase was 0.2 mL / min; the detection wavelength was 268 nm; and the injection volume was 1 μL.

5. The method according to claim 1, characterized in that The similarity of UPLC characteristic spectra of honey bran fried Codonopsis pilosula (Dongshen) slices, standard decoction and its formula granules was evaluated by using a traditional Chinese medicine chromatographic fingerprint similarity evaluation system, and a UPLC standard characteristic spectrum of honey bran fried Codonopsis pilosula (Dongshen) slices, standard decoction and its formula granules consisting of 9 characteristic peaks was obtained, wherein Peak 7 (S): Codonoside I; Peak 8: Codonopsis acetylene glycoside nin; Peak 9: Codonopsis acetylene glycoside; in the honey bran fried Codonopsis pilosula (Dongshen) raw material characteristic spectrum, Codonopsis acetylene glycoside I peak was used as the reference peak S peak, and the relative retention time of each characteristic peak and the S peak was calculated, and the relative retention time was within ±10% of the specified value, and the specified values ​​were: 0.23 (peak 1), 0.26 (peak 2), 0.31 (peak 3), 0.35 (peak 4), 0.42 (peak 5), 0.81 (peak 6), 1.14 (peak 8), and 1.27 (peak 9), respectively.

6. The method according to claim 1, characterized in that The similarity of the UPLC characteristic spectrum of Codonopsis pilosula (Dongshen) medicinal material was evaluated by using the similarity evaluation system of traditional Chinese medicine chromatographic fingerprint, and a UPLC standard characteristic spectrum of Codonopsis pilosula (Dongshen) medicinal material consisting of 10 characteristic peaks was obtained, wherein Peak 7 (S): Codonoside I; Peak 8: Codonopsis arginine; Peak 9: Codonopsis arginine; In the characteristic spectrum of Codonopsis pilosula (Dongshen), the Codonopsis arginine I peak was used as the reference peak S peak, and the relative retention time of each characteristic peak and the S peak was calculated, and the relative retention time was within ±10% of the specified value, and the specified values ​​were: the specified values ​​were: 0.23 (peak 1), 0.26 (peak 2), 0.31 (peak 3), 0.35 (peak 4), 0.42 (peak 5), 0.63 (peak 6), 0.81 (peak 7), 1.14 (peak 9), and 1.27 (peak 10).

7. The method according to claim 1, characterized in that When the raw material of honey bran fried Codonopsis (Codonopsis pilosula) is honey bran fried Codonopsis pilosula slices, the pretreatment is: add water to the honey bran fried Codonopsis pilosula (Codonopsis pilosula) slices for decoction, filter, add water to the residue for ultrasonic treatment, cool, shake well, filter, and obtain; the water decoction time is 20 to 30 minutes; the ultrasonic power is 600w, the frequency is 40KHz, and the ultrasonic time is 20 to 30 minutes.

8. The method according to claim 1, characterized in that When the raw material of honey bran fried Codonopsis (Codonopsis pilosula) is honey bran fried Codonopsis pilosula (Codonopsis pilosula) standard decoction or formula granules, the pretreatment is: add water to the honey bran fried Codonopsis pilosula (Codonopsis pilosula) standard decoction or formula granules for ultrasonic treatment, cool, shake well, and filter to obtain; the ultrasonic power is 600w, the frequency is 40KHz, and the ultrasonic time is 20 to 30min; the mass volume ratio of the honey bran fried Codonopsis pilosula (Codonopsis pilosula) standard decoction or formula granules and water is 2g:25mL.

9. The method according to claim 1, characterized in that: The time for decocting the Codonopsis pilosula (Dangshen) medicinal material in water is 20 to 30 minutes; the ultrasonic power is 600w, the frequency is 40KHz, and the ultrasonic time is 20 to 30 minutes.

10. A method for identifying honey bran fried Codonopsis pilosula (Dangshen) decoction pieces, standard decoctions, formula granules and Codonopsis pilosula (Dangshen) medicinal materials, characterized in that, The identification is performed by the method described in any one of claims 1 to 9; the UPLC characteristic spectrum of Codonopsis pilosula (Dongshen) medicinal material has 10 peaks; the UPLC characteristic spectrum of Codonopsis pilosula (Dongshen) decoction pieces and its standard decoction and formula granules fried with honey and bran has 9 peaks; Codonopsis pilosula (Dongshen) medicinal material has 6 more peaks than the raw material of Codonopsis pilosula (Dongshen) fried with honey and bran, and the two can be identified.

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