A method for constructing the fingerprint spectrum of Shenfushu granules

Abstract

This invention belongs to the field of pharmaceutical analysis, specifically relating to a method for constructing a fingerprint spectrum of Shenfushu granules. The method for constructing a fingerprint spectrum of Shenfushu granules provided by this invention features high accuracy, high stability, good repeatability, good specificity, and strong specificity. The fingerprint spectrum analysis method for Shenfushu granules, a traditional Chinese medicine preparation for treating chronic renal failure, established by this invention can not only qualitatively analyze and identify the seven chemical components contained in this preparation, but also simultaneously present the characteristic peaks of five traditional Chinese medicines in the prescription, achieving the goal of overall quality control of multiple medicinal ingredients.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical analysis, specifically relating to a method for constructing a fingerprint spectrum of Shenfushu granules. Background Technology

[0002] Shenfushu granules are composed of traditional Chinese medicines such as rhubarb, immature bitter orange peel, salvia miltiorrhiza, and licorice. This formula has the functions of clearing the bowels and removing turbidity, promoting blood circulation and removing blood stasis. It is used for chronic renal failure, azotemia, and early-stage uremia, specifically those with symptoms of turbidity and blood stasis. This traditional Chinese medicine preparation is a medical institution preparation independently developed by the Second Xiangya Hospital of Central South University, and has a clinical use history of over 30 years with significant clinical efficacy.

[0003] However, traditional Chinese medicine (TCM) compound preparations have complex components and require careful compatibility in their formulation. The synergistic effect of all the herbs in the formula is crucial for treating a specific disease. Therefore, evaluating drug quality solely based on the content of a single component has limitations. Currently, quality testing methods for TCM compound preparations can only detect one or a few components, failing to comprehensively characterize the components of TCM. In particular, the marker components in this preparation include salvianolic acid B, naringin, and glycyrrhizic acid, and interference exists between the detection peaks of these various marker components.

[0004] Therefore, determining which marker components to use for formulation quality monitoring, and how to ensure that all marker components can be effectively detected during the testing process, have become technical problems that urgently need to be solved by those skilled in the art. Summary of the Invention

[0005] In view of this, the first objective of the present invention is to address the problems existing in the prior art by providing a characteristic spectrum of traditional Chinese medicine compound preparations, wherein the spectrum can comprehensively reflect the components of traditional Chinese medicine compound preparations and realize the quality control of traditional Chinese medicine preparations.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] A method for constructing a fingerprint spectrum of Shenfushu granules, comprising the following steps:

[0008] I) Preparation of single and mixed reference solutions: Weigh appropriate amounts of glycyrrhizin, salvianolic acid B, glycyrrhizic acid ammonium, astilbene, naringin, neohesperidin, codonopsis glycoside, aloe-emodin, rhein, emodin, chrysophanol, emodin methyl ether, and tanshinone IIA reference standards, dissolve and dilute with methanol to prepare single reference solutions, and mix appropriate amounts of single reference solutions to prepare mixed reference solutions;

[0009] II) Preparation of test solution: Take 0.5g of Shenfushu granules, crush them, weigh them accurately, put them in a stoppered conical flask, add 25mL of 75% methanol accurately, stopper tightly, weigh them, sonicate for 20min, let stand to room temperature, weigh them again, make up the lost weight with 75% methanol, shake well, filter, and take the filtrate.

[0010] III) Preparation of solutions for each herb: Take 0.5g of each of the following powders: Salvia miltiorrhiza, rhubarb, Smilax glabra, Polygonatum sibiricum, Codonopsis pilosula, Citrus aurantium, Ostrea gigas, and Glycyrrhiza uralensis. Weigh them accurately and place them in a stoppered conical flask. Add 25mL of 75% methanol accurately, stopper tightly, weigh, sonicate for 20min, let stand to room temperature, weigh again, replenish the lost weight with methanol, shake well, filter, and collect the filtrate.

[0011] IV) High-performance liquid chromatography (HPLC) analysis: Accurately pipette equal volumes of the single reference solution, mixed reference solution, test solution, and each medicinal material solution, and inject them separately into the HPLC instrument for analysis to obtain the chromatograms of each standard solution and reference solution. The chromatographic conditions are as follows:

[0012] Using octadecylsilane-bonded silica gel as the packing material; using acetonitrile as mobile phase A and 0.1% phosphoric acid solution as mobile phase B, gradient elution was performed according to the specifications in the table below; the detection wavelength was 260 nm; the column temperature was 30℃-35℃; the flow rate was 0.9-1.1 mL / min; the theoretical plate number calculated based on the Danshensu B peak should not be less than 3000;

[0013]

[0014] V) Construction of fingerprint spectrum: Based on the injection results of single reference solution, mixed reference solution, and solutions of various medicinal materials, the retention times of chromatographic peaks were compared to obtain the fingerprint spectrum of Shenfushu granules. The fingerprint spectrum is labeled with 17 chromatographic peaks in chronological order and numbered 1 to 17, including qualitative peaks of 7 chemical components and characteristic peaks of 5 traditional Chinese medicines. Among them, peak 5 is naringin, peak 6 is neohesperidin, peak 8 is salvianolic acid B, peak 13 is aloe-emodin, peak 14 is glycyrrhizic acid, peak 15 is rhein, and peak 17 is emodin. Peaks 1, 2, and 3 are specifically associated with Polygonatum sibiricum, peaks 4, 5, and 6 are specifically associated with Citrus aurantium, peaks 7, 8, and 9 are specifically associated with Salvia miltiorrhiza, peaks 10, 13, 16, and 17 are associated with Rheum palmatum, peaks 11, 12, and 14 are specifically associated with Glycyrrhiza uralensis, and peak 15 is shared by both Rheum palmatum and Citrus aurantium.

[0015] It is worth noting that naringin and neohesperidin are indicator components of Citrus aurantium, tanshinone B is an indicator component of Salvia miltiorrhiza, glycyrrhizic acid is an indicator component of Glycyrrhiza uralensis, and aloe-emodin, rhein, and emodin are indicator components of Rheum palmatum. Furthermore, the peak assignments in the fingerprint spectrum of the aforementioned Shenfushu granules are shown in Table 2.

[0016] Table 2. Peak assignments in the fingerprint spectrum of Shenfushu granules.

[0017]

[0018]

[0019] It is particularly noteworthy that fingerprinting technology, as a comprehensive and quantifiable analytical tool for the quality control of traditional Chinese medicine (TCM), is widely used. It is characterized by its large information content and strong characteristic features, effectively, comprehensively, and scientifically reflecting the authenticity, stability, and uniformity of the quality of TCM materials and preparations. Meanwhile, high-performance liquid chromatography (HPLC) features high separation efficiency, fast analysis speed, high detection sensitivity, and wide application range. Unaffected by sample volatility and thermal stability, most TCM components can be analyzed and detected using HPLC. Therefore, HPLC has become the preferred method for constructing fingerprint profiles of TCM.

[0020] Specifically, the compound preparations detected in this invention include ingredients such as Citrus aurantium, Salvia miltiorrhiza, and Glycyrrhiza uralensis. The detection peaks of the marker components of these three traditional Chinese medicinal materials—tanshinone B, naringin, and glycyrrhizic acid—directly interfere with each other, making it difficult to construct the fingerprint spectrum of the compound preparation. This invention, by adjusting the gradient ratio of the mobile phase, specifically and exclusively distinguishes the detection peaks of tanshinone B, naringin, and glycyrrhizic acid, separating the three target peaks from their neighboring peaks with a resolution greater than 1.

[0021] Furthermore, using peak 8 (tanshinone B) as the reference peak and a relative retention time of 1, the average relative retention times of the chromatographic peaks 1 to 17 were calculated to be: 0.16, 0.20, 0.21, 0.74, 0.78, 0.87, 0.91, 1.00, 1.10, 1.14, 1.47, 1.55, 1.60, 1.62, 1.69, 1.78, and 1.93.

[0022] Furthermore, the conditions for ultrasonic treatment in steps II) and III) are 300W and 40kHz.

[0023] Furthermore, in step III), the column temperature is 30°C and the flow rate is 1.0 mL / min.

[0024] Compared with existing technologies, the fingerprint analysis method for Shenfushu granules, a traditional Chinese medicine preparation for treating chronic renal failure, established in this invention can not only perform qualitative analysis and identification of the seven chemical components contained in this preparation, but also simultaneously present the characteristic peaks of the five traditional Chinese medicines in the prescription of this preparation, thereby achieving the purpose of overall quality control of multiple medicinal ingredients. Attached Figure Description

[0025] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are merely embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0026] Figure 1 This is the chromatogram of the mixed reference standard obtained in Example 4 of the present invention.

[0027] Figure 2 This is the fingerprint spectrum of Shenfushu granules obtained in Example 4 of the present invention.

[0028] Figure 3 This is a chromatogram of the test sample under different mobile phase gradient elution programs as measured in Comparative Example 1 of this invention.

[0029] Figure 4 This is a chromatogram of the test sample at different detection wavelengths obtained in Comparative Example 2 of the present invention. Detailed Implementation

[0030] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0031] The term "embodiment" used herein, as an example, is not necessarily to be construed as superior to or better than other embodiments. Performance testing in the embodiments of this application, unless otherwise specified, employs conventional testing methods in the art. It should be understood that the terminology used in this application is merely for describing particular implementations and is not intended to limit the scope of this disclosure.

[0032] Unless otherwise stated, the technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains; other experimental methods and technical means not specifically mentioned herein refer to experimental methods and technical means commonly used by one of ordinary skill in the art.

[0033] To better illustrate the content of this application, numerous specific details are provided in the following detailed embodiments. Those skilled in the art should understand that this application can be implemented even without certain specific details. In the embodiments, some methods, means, instruments, and devices well-known to those skilled in the art are not described in detail in order to highlight the main points of this application. Without conflict, the technical features disclosed in the embodiments of this application can be arbitrarily combined, and the resulting technical solutions belong to the content disclosed in the embodiments of this application.

[0034] To better understand the present invention, the following embodiments are provided for further detailed description of the present invention, but they should not be construed as limiting the present invention. Any non-essential improvements and adjustments made by those skilled in the art based on the above-described invention are also considered to fall within the protection scope of the present invention.

[0035] The instruments, equipment, and reagents used in these embodiments of the invention are as follows:

[0036] Instruments and equipment:

[0037] Agilent 1260 Infinity II high-performance liquid chromatograph (Agilent Technologies), 0.01% balance (SQP Quintix 224-1CN, Sartorius Scientific Instruments Co., Ltd.), ultrasonic cleaner (SB-5200D, Ningbo Xinzhi Biotechnology Co., Ltd.).

[0038] Reference standards, test samples, and processed medicinal materials:

[0039] Shenfushu Granules (batch numbers 230601, 230602, 230603)

[0040] Salvia miltiorrhiza (22010810, Origin: Shandong)

[0041] Rhubarb (22010705, Origin: Qinghai)

[0042] Smilax glabra (22022808, origin: Guangdong)

[0043] Polygonatum (22021008, Origin: Hunan)

[0044] Codonopsis pilosula (22032608, origin: Gansu)

[0045] Citrus aurantium (22012004, origin: Hunan)

[0046] Oyster (22122411, Origin: Guangdong)

[0047] Licorice (22021001, Origin: Inner Mongolia)

[0048] Glycyrrhizin (111610-202209, China National Institutes for Food and Drug Control)

[0049] Tanshinone B (111562-201917, China National Institutes for Food and Drug Control)

[0050] Astilbin (111798-201805, China National Institutes for Food and Drug Control)

[0051] Naringin (110722-202116, China National Institutes for Food and Drug Control)

[0052] Neohesperidin (111857-201804, China National Institutes for Food and Drug Control)

[0053] Codonopsis glycosides (111732-201908, China National Institutes for Food and Drug Control)

[0054] Ammonium glycyrrhizate (110731-202122, China National Institutes for Food and Drug Control) and aloe-emodin (110795-202011, China National Institutes for Food and Drug Control)

[0055] Rhein (110757-201607, China National Institutes for Food and Drug Control)

[0056] Emodin (110756-201913, China National Institutes for Food and Drug Control)

[0057] Rhein (110796-201922, China National Institutes for Food and Drug Control)

[0058] Emodin methyl ether (110758-202218, China National Institutes for Food and Drug Control)

[0059] Tanshinone IIA (110766-202022, China National Institutes for Food and Drug Control)

[0060] Example 1

[0061] Precision test

[0062] Take 0.5g of Shenfushu granules, crush them, weigh them accurately, place them in a stoppered conical flask, add 25mL of methanol accurately, stopper tightly, weigh them, sonicate for 20min, let stand to room temperature, weigh them again, make up the lost weight with methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0063] Take the same sample solution of this traditional Chinese medicine preparation, inject it six times consecutively under the chromatographic conditions of this invention, and record the chromatograms. Using tanshinone B (peak 8) as the reference peak, calculate the RSD of the relative retention time of each chromatographic peak, which is 0.01% to 0.16%, and the RSD of the relative peak area, which is 0.095% to 5.24%, indicating that the instrument precision is good.

[0064] Table 3. Results of relative retention time in precision tests

[0065] # First injection 2nd injection 3rd injection 4th injection 5th stitch 6th stitch average value RSD / % 1 0.1593 0.1597 0.1598 0.1598 0.1599 0.1600 0.1597 0.16 2 0.1983 0.1989 0.1989 0.1990 0.1989 0.1991 0.1989 0.14 3 0.2081 0.2085 0.2086 0.2087 0.2086 0.2089 0.2086 0.13 4 0.7357 0.7366 0.7366 0.7363 0.7365 0.7368 0.7364 0.05 5 0.7832 0.7839 0.7840 0.7837 0.7838 0.7841 0.7838 0.04 6 0.8664 0.8469 0.8669 0.8666 0.8666 0.8667 0.8634 0.93 7 0.9119 0.9122 0.9121 0.9120 0.9120 0.9121 0.9121 0.01 8 1.0000 1.0000 1.0000 1.0000 1.0000 1.0000 1.0000 0.00 9 1.1041 1.1036 1.1038 1.1037 1.1038 1.1038 1.1038 0.02 10 1.1386 1.1374 1.1372 1.1371 1.1369 1.1371 1.1374 0.06 11 1.4711 1.4698 1.4690 1.4694 1.4693 1.4697 1.4697 0.05 12 1.5511 1.5498 1.5488 1.5493 1.5492 1.5497 1.5497 0.05 13 1.5951 1.5937 1.5929 1.5931 1.5931 1.5937 1.5936 0.05 14 1.6157 1.6141 1.6131 1.6136 1.6136 1.6141 1.6140 0.06 15 1.6860 1.6835 1.6824 1.6826 1.6824 1.6832 1.6833 0.08 16 1.7751 1.7733 1.7723 1.7730 1.7731 1.7737 1.7734 0.05 17 1.9328 1.9307 1.9295 1.9302 1.9302 1.9310 1.9308 0.06

[0066] Table 4. Results of relative peak area in precision test

[0067]

[0068]

[0069] Example 2

[0070] Stability test

[0071] Take 0.5g of Shenfushu granules, crush them, weigh them accurately, place them in a stoppered conical flask, add 25mL of methanol accurately, stopper tightly, weigh them, sonicate for 20min, let stand to room temperature, weigh them again, make up the lost weight with methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0072] The samples were placed at room temperature for 0, 12, 24, 36, 48, 60, and 72 hours and then measured. Using salvianolic acid B (peak 8) as the reference peak, the RSD of the relative retention time for each common peak was calculated to be 0.01%–0.19%, and the RSD of the relative peak area for the remaining 16 peaks (excluding peak 15) was 0.66%–5.30%. This indicates that the sample is stable within 72 hours at room temperature.

[0073] Table 5. Results of relative retention times in stability tests

[0074]

[0075]

[0076] Table 6. Results of relative peak area in stability tests

[0077]

[0078] Example 3

[0079] Repeatable experiments

[0080] Take 0.5g of Shenfushu granules, crush them, weigh them accurately, place them in a stoppered conical flask, add 25mL of methanol accurately, stopper tightly, weigh them, sonicate for 20min, let stand to room temperature, weigh them again, make up the lost weight with methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0081] Six sample solutions were prepared in parallel and injected separately. Using salvianolic acid B (peak 8) as the reference peak, the RSD of the relative retention time of each common peak was calculated to be 0.07%–0.75%, and the RSD of the relative peak area of ​​the remaining 16 peaks (excluding peak 12) was 0.40%–4.80%, indicating that the method has good repeatability.

[0082] Table 7. Results of Relative Retention Times in Repeatability Tests

[0083]

[0084] Table 8. Results of Relative Peak Area in Repeatability Tests

[0085]

[0086]

[0087] The above experimental results show that the fingerprint detection method for Shenfushu granules established in this invention has good precision, stability and repeatability, and can effectively characterize the quality of this traditional Chinese medicine preparation, which is beneficial for comprehensive quality monitoring.

[0088] Example 4

[0089] Establishment of characteristic spectra of traditional Chinese medicine compound preparations

[0090] I) Preparation of single and mixed reference solutions: Weigh appropriate amounts of glycyrrhizin, salvianolic acid B, glycyrrhizic acid ammonium, astilbene, naringin, neohesperidin, codonopsis glycoside, glycyrrhizic acid, aloe-emodin, rhein, emodin, chrysophanol, emodin methyl ether, and tanshinone IIA reference standards, dissolve and dilute with methanol to prepare single reference solutions, and mix appropriate amounts of single reference solutions to prepare mixed reference solutions;

[0091] II) Preparation of test solution: Take 0.5g of Shenfushu granules, crush them, weigh them accurately, put them in a stoppered conical flask, add 25mL of methanol accurately, stopper tightly, weigh them, sonicate for 20min, let stand to room temperature, weigh them again, make up the lost weight with methanol, shake well, filter, and take the filtrate.

[0092] III) Preparation of solutions for each herb: Take 0.5g each of Salvia miltiorrhiza, rhubarb, Smilax glabra, Polygonatum sibiricum, Codonopsis pilosula, Citrus aurantium, Ostrea gigas, and Glycyrrhiza uralensis, crush them, weigh them accurately, place them in a stoppered conical flask, add 25mL of methanol accurately, stopper tightly, weigh them, sonicate for 20min, let stand to room temperature, weigh them again, make up the lost weight with methanol, shake well, filter, and take the filtrate.

[0093] IV) High-performance liquid chromatography (HPLC) analysis: Accurately pipette equal volumes of the single reference solution, mixed reference solution, test solution, and each medicinal material solution, and inject them separately into the HPLC instrument for analysis to obtain the chromatograms of each standard solution and reference solution. The chromatographic conditions are as follows:

[0094] Using octadecylsilane-bonded silica gel as the packing material; using acetonitrile as mobile phase A and 0.1% phosphoric acid solution as mobile phase B, gradient elution was performed according to the specifications in the table below; the detection wavelength was 286 nm; the column temperature was 30 °C; the flow rate was 1.0 mL / min; the theoretical plate number calculated based on the Danshensu B peak should not be less than 3000;

[0095]

[0096] V) Construction of fingerprint spectrum: Based on the injection results of single reference solution, mixed reference solution, and solutions of various medicinal materials, the retention times of chromatographic peaks were compared to obtain the fingerprint spectrum of Shenfushu granules. The fingerprint spectrum is labeled with 17 chromatographic peaks in chronological order and numbered 1 to 17, including qualitative peaks of 7 chemical components and characteristic peaks of 5 traditional Chinese medicines. Among them, peak 5 is naringin, peak 6 is neohesperidin, peak 8 is salvianolic acid B, peak 13 is aloe-emodin, peak 14 is glycyrrhizic acid, peak 15 is rhein, and peak 17 is emodin. Peaks 1, 2, and 3 are specifically associated with Polygonatum sibiricum, peaks 4, 5, and 6 are specifically associated with Citrus aurantium, peaks 7, 8, and 9 are specifically associated with Salvia miltiorrhiza, peaks 10, 13, 16, and 17 are associated with Rheum palmatum, peaks 11, 12, and 14 are specifically associated with Glycyrrhiza uralensis, and peak 15 is shared by both Rheum palmatum and Citrus aurantium.

[0097] To further demonstrate the beneficial effects of the present invention and to better understand the present invention, the technical progress of the present invention is determined by the following comparative examples, but these should not be construed as limiting the present invention. Other measurement experiments conducted by those skilled in the art based on the above-described invention to obtain product properties and applications based on the above properties are also considered to fall within the protection scope of the present invention.

[0098] Comparative Example 1

[0099] The experiment was conducted according to the preparation and testing methods of the test solution in Example 4, the difference being that different mobile phase gradient elution programs as shown in Tables 9-10 were used. The chromatograms of the test sample from Example 4 were compared with the chromatograms of the test sample under different mobile phase gradient elution programs in Comparative Example 1. Figure 3Comparing a and b), the results show that the chromatographic peak separation effect of the mobile phase gradient elution program shown in Table 9-10 is not good.

[0100] Table 9. Mobile Phase Gradient Elution Procedure a

[0101]

[0102] Table 10 Mobile Phase Gradient Elution Program (b)

[0103]

[0104] Comparative Example 2

[0105] The test was conducted according to the preparation and testing methods of the test solution in Example 4, the difference being that a detection wavelength of 210–300 nm was used to determine the fingerprint spectrum of the Shenfushu granules. The results obtained ( Figure 4 a is 210nm. Figure 4 (b = 300nm) It can be seen that the intensity of each chromatographic peak is better when the detection wavelength is 260nm, so 260nm is selected as the detection wavelength.

[0106] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for constructing a fingerprint of Shenfushu granules, characterized in that, The steps are as follows: I) Preparation of single control solution and mixed control solution: take appropriate amount of glycyrrhizin, salvianolic acid B, ammonium glycyrrhizate, astilbin, naringin, neohesperidin, tenuifolin, aloe-emodin, rhein, emodin, chrysophanol, dan-shen ketone II A control, dissolve in methanol to constant volume to prepare single control solution, take appropriate amount of single control solution to mix to prepare mixed control solution; II) Preparation of test solution: take 0.5g of Shenfushu granules, crush, accurately weigh, place in a conical flask with a plug, accurately add 25mL of 75% methanol, tightly plug, weigh, ultrasonic treatment for 20min, stand to room temperature, weigh again, make up the weight loss with 75% methanol, shake well, filter, take the filtrate; III) Preparation of each medicinal material solution: take 0.5g of salvia miltiorrhiza, rhubarb, smilax glabra, polygonatum, codonopsis pilosula, aurantium, oyster, licorice powder, accurately weigh, place in a conical flask with a plug, accurately add 25mL of 75% methanol, tightly plug, weigh, ultrasonic treatment for 20min, stand to room temperature, weigh again, make up the weight loss with methanol, shake well, filter, take the filtrate; IV) High performance liquid chromatography analysis: accurately take equal amount of single control solution, mixed control solution, test solution and each medicinal material solution, inject into liquid chromatograph for determination, to obtain chromatogram of each standard solution and reference solution, chromatographic conditions are as follows: Octadecylsilane bonded silica gel as filler; Acetonitrile as mobile phase A, 0.1% phosphoric acid solution as mobile phase B, gradient elution according to the following table; detection wavelength is 260nm; column temperature is 30-35℃; flow rate is 0.9-1.1mL / min; theoretical plate number calculated by salvianolic acid B peak should not be less than 3000; V) Fingerprint construction: according to injection results of single control solution, mixed control solution and each medicinal material solution, compare chromatographic peak retention time, to develop the fingerprint of Shenfushu granules; the fingerprint is marked with 17 chromatographic peaks in time sequence and numbered 1-17, including 7 kinds of chemical components and 5 kinds of characteristic peaks of traditional Chinese medicine; among them, peak 5 is naringin, peak 6 is neohesperidin, peak 8 is salvianolic acid B, peak 13 is aloe-emodin, peak 14 is glycyrrhizic acid, peak 15 is rhein, peak 17 is emodin; peaks 1, 2 and 3 are specific to polygonatum, peaks 4, 5 and 6 are specific to aurantium, peaks 7, 8 and 9 are specific to salvia miltiorrhiza, peaks 10, 13, 16 and 17 belong to rhubarb, peaks 11, 12 and 14 belong to licorice, and peak 15 is common to rhubarb and aurantium.

2. The method for constructing the fingerprint spectrum of Shenfushu granules according to claim 1, characterized in that, Take peak 8 salvianolic acid B as reference peak, relative retention time is 1, calculate the average relative retention time of peaks 1-17, which are 0.16, 0.20, 0.21, 0.74, 0.78, 0.87, 0.91, 1.00, 1.10, 1.14, 1.47, 1.55, 1.60, 1.62, 1.69, 1.78 and 1.93 respectively.

3. The method for constructing the fingerprint spectrum of Shenfushu granules according to claim 1, characterized in that, The ultrasonic treatment conditions in steps II) and III) are 300W, 40kHZ.

4. The method for constructing the fingerprint spectrum of Shenfushu granules according to claim 1, characterized in that, The column temperature in step III) is 30 °C; the flow rate is 1.0 mL / min. The column temperature in step III) is 30 °C; the flow rate is 1.0 mL / min.

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