Construction method of fingerprint spectrum of Shenxiong Tongmai capsule

Abstract

The invention relates to a construction method of a fingerprint spectrum of Shenxiong Tongmai capsule, which comprises the following steps: dissolving a ferulic acid standard substance, a hesperidin standard substance, a ginsenoside Re standard substance, a neohesperidin standard substance and a naringin standard substance in methanol to prepare a mixed reference substance solution; the method comprises the following steps: respectively dissolving contents of Shenxiong Tongmai capsules in a plurality of batches in a methanol aqueous solution to prepare a plurality of test solutions; respectively carrying out high performance liquid chromatography detection on the mixed reference substance solution and the plurality of test sample solutions to obtain a high performance liquid chromatogram of the mixed reference substance solution and high performance liquid chromatograms of the plurality of test sample solutions; determining characteristic peaks and identifying chemical components of the characteristic peaks, and calculating relative retention time of other characteristic peaks by taking the characteristic peak corresponding to hesperidin as a reference peak to obtain the fingerprint spectrum. The fingerprint spectrum obtained by the construction method disclosed by the invention can well reflect information of a plurality of index components of the Shenxiong Tongmai capsule.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical analysis, and particularly relates to a method for constructing a fingerprint of Shenxiong Tongmai Capsule. Background Art

[0002] Angina pectoris belongs to a clinical syndrome, the main feature of which is chest pain caused by temporary myocardial ischemia, and it is the most common manifestation of coronary heart disease. It usually appears in patients with at least 50% stenosis in the lumen diameter of at least one main branch of the coronary artery. When the patient is physically or mentally stressed, the coronary blood flow cannot meet the myocardial metabolic needs, thereby leading to myocardial ischemia and triggering an angina attack. In the scope of traditional Chinese medicine, angina pectoris mainly corresponds to the syndrome of "chest impediment and heart pain", and its pathogenesis is qi deficiency of the heart and obstruction of the heart vessels; the pathological changes mainly present a state of deficiency in origin and excess in superficiality, with deficiency and excess intermingled, among which qi deficiency and blood stasis are the most common. Therefore, the treatment should mainly focus on supplementing qi and activating blood circulation, dredging the meridians and relieving pain, so as to make the qi of the heart strong, the blood circulation powerful, the stasis eliminated, and the meridians unobstructed, and thus all symptoms can be cured.

[0003] Shenxiong Tongmai Capsule is composed of borneol, chuanxiong rhizome, earthworm, cassia twig, red ginseng, acorus tatarinowii rhizome, Corydalis yanhusuo, Gleditsia sinensis thorn, and immature bitter orange. This is a clinical prescription obtained based on the essence of the pathogenesis of qi deficiency and blood stasis in coronary heart disease angina pectoris and combined with long-term clinical practice. This prescription pays attention to treating both the symptoms and the root causes, combining the main symptoms with the concurrent symptoms, and puts forward three principles for treating the root cause: supplementing qi of the heart, nourishing heart blood, and warming and consolidating heart yang; two principles for treating the superficiality: relieving chest impediment, dispelling dampness and resolving phlegm, promoting blood circulation to remove stasis and dredging the meridians; four auxiliary principles: relieving fullness in the chest and regulating the lung, soothing the liver and regulating qi, strengthening the spleen and harmonizing the stomach, and promoting the smooth passage of urine and feces. Shenxiong Tongmai Capsule has the effects of supplementing qi and activating blood circulation, dredging collaterals and relieving pain, and is clinically used for coronary heart disease and angina pectoris, especially suitable for patients with qi stagnation and blood stasis. It can be flexibly applied in the treatment of heart failure, hypotensive syndrome, arrhythmia, and the recovery period of myocardial infarction in combination with western medicine treatment plans, and has achieved remarkable results in clinical applications.

[0004] Hospital preparations are a hot topic in the research of new traditional Chinese medicines due to their rich theories and human use experience. Combining traditional Chinese medicine theories with modern technologies can produce traditional Chinese medicine prescriptions that restore the records in ancient medical books and meet modern clinical needs, effectively filling the gap in the use of drugs for difficult and complicated diseases and promoting the upgrading and innovation of the traditional Chinese medicine industry. Since the components of traditional Chinese medicine compounds are complex and the content differences are large, relying solely on qualitative and quantitative detection methods for single or several components cannot comprehensively detect and control the quality of the preparation finished products, and it is difficult to ensure the reproducibility of the pharmacological effects.

[0005] In order to better control the quality of drugs and ensure clinical efficacy, it is of great practical significance to establish a finished product fingerprint method that contains multi-component information, and it can also lay a solid foundation for subsequent research. Summary of the Invention

[0006] The object of the present invention is to provide a method for constructing a fingerprint of Shenxiong Tongmai Capsules in view of the deficiencies in the prior art.

[0007] To achieve the above object, the technical solution adopted by the present invention is:

[0008] Provide a method for constructing a fingerprint of Shenxiong Tongmai Capsules, wherein the Shenxiong Tongmai Capsules are composed of borneol, chuanxiong rhizome, earthworm, cassia twig, red ginseng, acorus tatarinowii, rhizoma corydalis, Chinese honey locust spine, and fructus aurantii immaturus; the steps of the construction method include:

[0009] S1. Dissolve ferulic acid reference substance, hesperidin reference substance, ginsenoside Re reference substance, neohesperidin reference substance, and naringin reference substance in methanol to prepare a mixed reference substance solution;

[0010] Dissolve the contents of several batches of the Shenxiong Tongmai Capsules in a methanol aqueous solution to prepare several test solution;

[0011] S2. Perform high performance liquid chromatography (HPLC) detection on the mixed reference substance solution and several test solutions respectively to obtain the HPLC chromatogram of the mixed reference substance solution and the HPLC chromatograms of several test solutions;

[0012] Among them, the conditions for the HPLC detection include:

[0013] The column length of the chromatographic column is 150 mm - 300 mm; the inner diameter of the chromatographic column is 4 mm - 7 mm; the packing material of the chromatographic column is octadecylsilane chemically bonded silica gel, and the particle size of the octadecylsilane chemically bonded silica gel is 3.0 μm - 6.0 μm;

[0014] The detection wavelength is 195 nm - 230 nm; the detection temperature is 25 °C - 30 °C; the injection volume is 5 μL - 15 μL;

[0015] Mobile phase A is methanol, and mobile phase B is an aqueous phosphoric acid solution with a volume fraction of 0.1%; the flow rate is 0.80 mL / min - 1.20 mL / min; the elution method is gradient elution; by volume fraction, the gradient elution program includes:

[0016] At 0 min, mobile phase A is 5% and mobile phase B is 95%;

[0017] At 10 min, mobile phase A is 20% and mobile phase B is 80%;

[0018] At 15 min, mobile phase A is 40% and mobile phase B is 60%;

[0019] At 30 min, mobile phase A is 40% and mobile phase B is 60%;

[0020] At 50 min, mobile phase A is 75% and mobile phase B is 25%;

[0021] At 65 min, mobile phase A is 75% and mobile phase B is 25%;

[0022] At 70 min, mobile phase A is 100% and mobile phase B is 0%;

[0023] At 75 min, mobile phase A is 100% and mobile phase B is 0%;

[0024] S3. Determine the characteristic peaks in the high performance liquid chromatography (HPLC) chromatograms of several test sample solutions, and identify the chemical components of the characteristic peaks according to the HPLC chromatogram of the mixed reference substance solution;

[0025] Using the characteristic peak corresponding to hesperidin as the reference peak, calculate the relative retention time of other characteristic peaks, and thus obtain the fingerprint of the Shenxiong Tongmai Capsules.

[0026] Preferably, in the mixed reference substance solution, the mass-to-volume ratio of ferulic acid reference substance to methanol is 30 μg / mL - 60 μg / mL, the mass-to-volume ratio of hesperidin reference substance to methanol is 30 μg / mL - 60 μg / mL, the mass-to-volume ratio of ginsenoside Re reference substance to methanol is 150 μg / mL - 300 μg / mL, the mass-to-volume ratio of neohesperidin reference substance to methanol is 30 μg / mL - 60 μg / mL, and the mass-to-volume ratio of naringin reference substance to methanol is 30 μg / mL - 60 μg / mL.

[0027] Preferably, after dissolving the contents of several batches of the Shenxiong Tongmai Capsules in the methanol aqueous solution, ultrasonic treatment and filtration treatment are successively carried out to prepare several test sample solutions; among them,

[0028] The conditions of the ultrasonic treatment include: ultrasonic power is 150 W - 450 W, ultrasonic frequency is 20 kHz - 60 kHz, and ultrasonic time is 15 min - 45 min.

[0029] Preferably, in the methanol aqueous solution, the volume percentage of methanol is 40% - 60%; in the test sample solution, the mass-to-volume ratio of the contents of the Shenxiong Tongmai Capsules to the methanol aqueous solution is 0.03 g / mL - 0.2 g / mL.

[0030] Preferably, the conditions of the HPLC detection further include: calculated according to the characteristic peak corresponding to the hesperidin reference substance, the theoretical plate number is not less than 3000.

[0031] Preferably, the fingerprint includes 14 characteristic peaks.

[0032] Preferably, the fingerprint includes: a third characteristic peak corresponding to ferulic acid, a fifth characteristic peak corresponding to naringin, a sixth characteristic peak corresponding to hesperidin, a seventh characteristic peak corresponding to neohesperidin, and a tenth characteristic peak corresponding to ginsenoside Re.

[0033] Preferably, taking the sixth characteristic peak corresponding to hesperidin as the reference peak, the relative retention time of the third characteristic peak corresponding to ferulic acid is 0.720 - 0.880, the relative retention time of the fifth characteristic peak corresponding to naringin is 0.837 - 1.023, the relative retention time of the seventh characteristic peak corresponding to neohesperidin is 0.954 - 1.166, and the relative retention time of the tenth characteristic peak corresponding to ginsenoside Re is 1.242 - 1.518.

[0034] Preferably, taking the sixth characteristic peak corresponding to hesperidin as the reference peak, the fingerprint further includes: a first characteristic peak with a relative retention time of 0.179 - 0.221, a second characteristic peak with a relative retention time of 0.333 - 0.407, a fourth characteristic peak with a relative retention time of 0.774 - 0.946, an eighth characteristic peak with a relative retention time of 1.179 - 1.441, a ninth characteristic peak with a relative retention time of 1.215 - 1.485, an eleventh characteristic peak with a relative retention time of 1.346 - 1.654, a twelfth characteristic peak with a relative retention time of 1.386 - 1.694, a thirteenth characteristic peak with a relative retention time of 1.467 - 1.793, and a fourteenth characteristic peak with a relative retention time of 1.521 - 1.859.

[0035] The present invention adopts the above technical solutions, and compared with the prior art, has the following technical effects:

[0036] The fingerprint obtained by the construction method of the present invention can well reflect the information of multiple index components of Shenxiong Tongmai Capsules. The characteristic peak baseline of the fingerprint of the present invention is stable, the retention time distribution is uniform, and the sensitivity and resolution are good. At the same time, the operation is fast, reliable, and the cost is low, which can effectively, stably, and quickly monitor the quality of Shenxiong Tongmai Capsules comprehensively, and ensure the stability and quality controllability of Shenxiong Tongmai Capsules. Description of the Drawings

[0037] Figure 1 For the results of specificity investigation; among them, Figure A is the high - performance liquid chromatography (HPLC) chromatogram of the blank solution, Figure B is the HPLC chromatogram of the test solution, and Figure C is the HPLC chromatogram of the mixed reference substance solution;

[0038] Figure 2 It is the characteristic superposition diagram of the HPLC chromatograms of three batches of Shenxiong Tongmai Capsules;

[0039] Figure 3 It is the fingerprint spectrum of Shenxiong Tongmai Capsule. Specific Embodiments

[0040] The specific embodiments of the present invention will be described in detail below.

[0041] Unless otherwise defined, the technical terms or scientific terms used in the claims and the specification shall have the ordinary meanings understood by those of ordinary skill in the technical field to which the present invention pertains.

[0042] The use of the word "comprising" or similar words in the specification and claims of this patent application for the present invention means that the items appearing before "comprising" cover the items listed after "comprising" or their equivalents, and do not exclude other items.

[0043] The numerical values mentioned in the present invention include all numerical values that increase one unit by one unit from low to high. Here, it is assumed that there is an interval of at least two units between any lower value and higher value. For example, if it is said that a component amount or a physical quantity ranges from 1 to 100, 10 to 90 is more preferable, and 20 to 80 is most preferable, it is intended to express that numerical values such as 5 to 95, 14 to 76, 23 to 67, 32 to 58, 41 to 49, etc. are all clearly listed in this specification; for numerical values less than 1, 0.0001, 0.001, 0.01 or 0.1 is considered a more appropriate unit. The foregoing examples are for illustrative purposes only. In fact, all numerical combinations between the lowest value and the highest value listed are considered to be clearly listed in this specification in a similar manner.

[0044] Examples

[0045] Instruments and Reagents

[0046] Instruments: Analytical balance (Mettler-Toledo, XS204), pure water instrument (Millipore), ) ultrasonic cleaner (Kunshan Ultrasonic Instruments Co., Ltd., KQ3200E), bench-top high-speed refrigerated centrifuge (Shanghai Lishen Scientific Instruments Co., Ltd., Neofuge15R), Agilent high performance liquid chromatograph (Agilent Technologies, Agilent1290);

[0047] Reagents: Methanol (Shanghai Xingke High Purity Solvents Co., Ltd., chromatographic grade), phosphoric acid (Shanghai Xingke High Purity Solvents Co., Ltd., chromatographic grade), ultrapure water was prepared by the pure water instrument;

[0048] Test drugs: Ferulic acid reference standard (National Institutes for Food and Drug Control, 110773-201915), Hesperidin reference standard (National Institutes for Food and Drug Control, 110721-202220), Ginsenoside Re reference standard (National Institutes for Food and Drug Control, 110754-201626), Neohesperidin reference standard (National Institutes for Food and Drug Control, 111857-201804), Naringin reference standard (National Institutes for Food and Drug Control, 110722-202116), Shenxiong Tongmai Capsules (Shanghai Traditional Chinese Medicine Pharmaceutical Technology Co., Ltd., batch numbers 2401, 2402, 2403).

[0049] High Performance Liquid Chromatography Detection Conditions

[0050] Chromatographic column (InertSustain AQ-C18, 4.6 mm × 250 mm, 5 μm), that is, the column length of the chromatographic column is 250 mm; the inner diameter of the chromatographic column is 2.6 mm; the packing material of the chromatographic column is octadecylsilyl-bonded silica gel, and the particle size of the octadecylsilyl-bonded silica gel is 5 μm;

[0051] The detection wavelength is 210 nm; the detection temperature is 30 °C; the injection volume is 5 μL;

[0052] Mobile phase A is methanol, and mobile phase B is an aqueous phosphoric acid solution with a volume fraction of 0.1%; the flow rate is 1.0 mL / min; the elution method is gradient elution; by volume fraction, the gradient elution program includes:

[0053] At 0 min, mobile phase A is 5% and mobile phase B is 95%;

[0054] At 10 min, mobile phase A is 20% and mobile phase B is 80%;

[0055] At 15 min, mobile phase A is 40% and mobile phase B is 60%;

[0056] At 30 min, mobile phase A is 40% and mobile phase B is 60%;

[0057] At 50 min, mobile phase A is 75% and mobile phase B is 25%;

[0058] At 65 min, mobile phase A is 75% and mobile phase B is 25%;

[0059] At 70 min, mobile phase A is 100% and mobile phase B is 0%;

[0060] At 75 min, mobile phase A is 100% and mobile phase B is 0%.

[0061] Sample Preparation

[0062] Blank solution: An aqueous methanol solution with a volume percentage of 50% was used as the blank solution;

[0063] Mixed reference solution: Standards of ferulic acid, hesperidin, ginsenoside Re, neohesperidin, and naringin were dissolved in methanol to prepare a mixed reference solution;

[0064] Among them, the mass-volume ratio of ferulic acid standard to methanol was 50 μg / mL, the mass-volume ratio of hesperidin standard to methanol was 50 μg / mL, the mass-volume ratio of ginsenoside Re standard to methanol was 200 μg / mL, the mass-volume ratio of neohesperidin standard to methanol was 50 μg / mL, and the mass-volume ratio of naringin standard to methanol was 50 μg / mL;

[0065] Test solution: The contents of 3 g of Shenxiong Tongmai capsules were dissolved in 25 mL of an aqueous methanol solution with a volume percentage of 50%, and then subjected to ultrasonic treatment (ultrasonic power: 300 W, ultrasonic frequency: 40 kHz, ultrasonic time: 30 min) and filtration to obtain the test solution.

[0066] Specificity Investigation

[0067] Using the Shenxiong Tongmai capsules with batch number 2401 to prepare the test solution, the blank solution, the mixed reference solution, and the test solution were respectively subjected to high performance liquid chromatography detection to obtain the high performance liquid chromatogram of the blank solution, the high performance liquid chromatogram of the mixed reference solution, and the high performance liquid chromatogram of the test solution, as Figure 1 shown;

[0068] At a detection wavelength of 210 nm, the high performance liquid chromatogram of the test solution showed 14 characteristic peaks, which had the same chromatographic peaks at the same retention time as those in the high performance liquid chromatogram of the mixed reference solution, and there was no negative interference, indicating good specificity.

[0069] Precision Investigation

[0070] Using the Shenxiong Tongmai capsules with batch number 2401 to prepare the test solution, the test solution was continuously subjected to high performance liquid chromatography detection six times. Taking the characteristic peak corresponding to hesperidin as the reference peak, the relative retention time and relative peak area of other characteristic peaks were calculated, and the RSD values were calculated. The results are shown in the following table:

[0071]

[0072] The RSD value of the relative retention time was less than 2.0%, and the RSD value of the relative peak area was less than 5.0%, indicating good precision.

[0073] Repeatability Investigation

[0074] Six samples of test solutions were prepared from Shenxiong Tongmai capsules with batch number 2401, and the six samples of test solutions were respectively detected by high performance liquid chromatography (HPLC). Using the characteristic peak corresponding to hesperidin as the reference peak, the relative retention time and relative peak area of other characteristic peaks were calculated, and the RSD values were calculated. The results are shown in the following table:

[0075]

[0076]

[0077] The RSD value of the relative retention time was less than 2.0%, and the RSD value of the relative peak area was less than 5.0%, indicating good repeatability.

[0078] Stability Investigation

[0079] A test solution was prepared from Shenxiong Tongmai capsules with batch number 2401, and the test solution was detected by HPLC at 0 h, 2 h, 4 h, 6 h, 8 h, 10 h, 12 h, and 24 h respectively. Using the characteristic peak corresponding to hesperidin as the reference peak, the relative retention time and relative peak area of other characteristic peaks were calculated, and the RSD values were calculated. The results are shown in the following table:

[0080]

[0081]

[0082] The RSD value of the relative retention time was less than 2.0%, and the RSD value of the relative peak area was less than 5.0%, indicating good stability within 24 hours.

[0083] Sample Determination

[0084] Test solutions were prepared from Shenxiong Tongmai capsules with batch numbers 2401, 2402, and 2403 respectively, and the test solutions were detected by HPLC. The characteristic superposition diagram of the HPLC chromatograms of the three batches of Shenxiong Tongmai capsules is as shown in Figure 2 Using the characteristic peak corresponding to hesperidin as the reference peak, the relative retention time and relative peak area of other characteristic peaks were calculated, and the RSD values were calculated. The results are shown in the following table:

[0085]

[0086]

[0087] Matched with the "Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicines", the reference chromatogram was generated by the median method, and the fingerprint of Shenxiong Tongmai capsules was established, as shown inFigure 3 as shown

[0088] In summary, the fingerprint obtained by the construction method of the present invention can well reflect the information of multiple index components of Shenxiong Tongmai Capsule. The characteristic peaks of the fingerprint of the present invention have a stable baseline, uniform retention time distribution, good sensitivity and resolution, and at the same time, the operation is fast, reliable and low-cost, which can effectively, stably and quickly monitor the quality of Shenxiong Tongmai Capsule in an all-round way, and ensure the stability and quality controllability of Shenxiong Tongmai Capsule.

[0089] The above are only the preferred embodiments of the present invention, and do not limit the implementation manners and protection scope of the present invention. For those skilled in the art, it should be realized that all the equivalent substitutions and obvious changes made by using the description and illustration content of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for constructing a fingerprint of Shenxiong Tongmai Capsule, wherein the Shenxiong Tongmai Capsule is composed of borneol, chuanxiong, earthworm, cassia twig, red ginseng, calamus, yanhusuo, gleditsia spinosa, and immature bitter orange; characterized in that: The steps of the construction method include: S1. Dissolving a ferulic acid standard, a hesperidin standard, a ginsenoside Re standard, a neohesperidin standard, and a naringin standard in methanol to prepare a mixed reference solution; Dissolving the contents of several batches of the Shenxiong Tongmai Capsules in methanol aqueous solution respectively to prepare several test solutions; S2, respectively performing high performance liquid chromatography on the mixed reference solution and the plurality of test solution to obtain a high performance liquid chromatogram of the mixed reference solution and a plurality of high performance liquid chromatograms of the test solution; Wherein, the conditions for the high performance liquid chromatography detection include: The column length of the chromatographic column is 150 mm-300 mm; the inner diameter of the chromatographic column is 4 mm-7 mm; the filler of the chromatographic column is octadecylsilane bonded silica gel, and the particle size of the octadecylsilane bonded silica gel is 3.0 μm-6.0 μm; The detection wavelength is 195nm-230nm; the detection temperature is 25℃-30℃; the injection volume is 5μL-15μL; The mobile phase A is methanol, and the mobile phase B is a phosphoric acid aqueous solution with a volume fraction of 0.1%; the flow rate is 0.80mL / min-1.20mL / min; the elution method is gradient elution; in terms of volume fraction, the gradient elution procedure includes: at 0min, the mobile phase A is 5%, and the mobile phase B is 95%; At 10 min, mobile phase A was 20% and mobile phase B was 80%; At 15 min, mobile phase A was 40% and mobile phase B was 60%; At 30 min, mobile phase A was 40% and mobile phase B was 60%; At 50 min, mobile phase A was 75% and mobile phase B was 25%; At 65 min, mobile phase A was 75% and mobile phase B was 25%; At 70 min, mobile phase A was 100% and mobile phase B was 0%; At 75 min, mobile phase A was 100% and mobile phase B was 0%; S3, determining characteristic peaks in the HPLC chromatograms of several of the test solution, and identifying the chemical components of the characteristic peaks according to the HPLC chromatogram of the mixed reference solution; The characteristic peak corresponding to hesperidin is used as a reference peak, and the relative retention times of other characteristic peaks are calculated to obtain the fingerprint spectrum of the Shenxiong Tongmai Capsule.

2. The construction method according to claim 1, characterized in that: In the mixed reference solution, the mass volume ratio of the ferulic acid standard to the methanol is 30 μg / mL-60 μg / mL, the mass volume ratio of the hesperidin standard to the methanol is 30 μg / mL-60 μg / mL, the mass volume ratio of the ginsenoside Re standard to the methanol is 150 μg / mL-300 μg / mL, the mass volume ratio of the neohesperidin standard to the methanol is 30 μg / mL-60 μg / mL, and the mass volume ratio of the naringin standard to the methanol is 30 μg / mL-60 μg / mL.

3. The construction method according to claim 1, characterized in that: After dissolving the contents of several batches of the Shenxiong Tongmai Capsules in the methanol aqueous solution, ultrasonic treatment and filtration treatment were performed in sequence to prepare several test sample solutions; wherein, The ultrasonic treatment conditions include: ultrasonic power of 150W-450W, ultrasonic frequency of 20kHz-60kHz, and ultrasonic time of 15min-45min.

4. The construction method according to claim 1 or 3, characterized in that: In the methanol aqueous solution, the volume percentage of methanol is 40%-60%; in the test solution, the mass volume ratio of the content of the Shenxiong Tongmai Capsule to the methanol aqueous solution is 0.03g / mL-0.2g / mL.

5. The construction method according to claim 1, characterized in that: The conditions for the high performance liquid chromatography detection also include: the number of theoretical plates calculated according to the characteristic peaks corresponding to the hesperidin standard is not less than 3000.

6. The construction method according to claim 1, characterized in that: The fingerprint spectrum includes 14 characteristic peaks.

7. The construction method according to claim 1 or 6, characterized in that: The fingerprint spectrum includes: a third characteristic peak corresponding to ferulic acid, a fifth characteristic peak corresponding to naringin, a sixth characteristic peak corresponding to hesperidin, a seventh characteristic peak corresponding to neohesperidin, and a tenth characteristic peak corresponding to ginsenoside Re.

8. The construction method according to claim 7, characterized in that: Taking the sixth characteristic peak corresponding to hesperidin as the reference peak, the relative retention time of the third characteristic peak corresponding to ferulic acid is 0.720-0.880, the relative retention time of the fifth characteristic peak corresponding to naringin is 0.837-1.023, the relative retention time of the seventh characteristic peak corresponding to neohesperidin is 0.954-1.166, and the relative retention time of the tenth characteristic peak corresponding to ginsenoside Re is 1.242-1.

518.

9. The construction method according to claim 7, characterized in that: Taking the sixth characteristic peak corresponding to hesperidin as the reference peak, the fingerprint spectrum also includes: a first characteristic peak with a relative retention time of 0.179-0.221, a second characteristic peak with a relative retention time of 0.333-0.407, a fourth characteristic peak with a relative retention time of 0.774-0.946, an eighth characteristic peak with a relative retention time of 1.179-1.441, a ninth characteristic peak with a relative retention time of 1.215-1.485, an eleventh characteristic peak with a relative retention time of 1.346-1.654, a twelfth characteristic peak with a relative retention time of 1.386-1.694, a thirteenth characteristic peak with a relative retention time of 1.467-1.793, and a fourteenth characteristic peak with a relative retention time of 1.521-1.859.

Images