Fingerprint spectrum and content determination method of heart-tonifying and nerve-soothing preparation for treating insomnia

Abstract

The invention provides a fingerprint spectrum and content determination method of a heart-tonifying and nerve-soothing preparation for treating insomnia, and belongs to the technical field of traditional Chinese medicines. On one hand, the invention provides a fingerprint spectrum construction method of the heart-tonifying and nerve-soothing preparation for treating insomnia; secondly, the invention provides a fingerprint spectrum of the heart-tonifying and nerve-soothing preparation for treating insomnia; in the third aspect, the invention provides application of the fingerprint spectrum of the heart-tonifying and nerve-soothing preparation for treating insomnia as a standard fingerprint spectrum in detection and quality control of the heart-tonifying and nerve-soothing preparation. The fingerprint spectrum can be used for simultaneously detecting various chemical components in the heart-tonifying and nerve-soothing preparation, is high in separation degree, and can systematically, stably and fully reflect the quality of a product, so that the quality of the heart-tonifying and nerve-soothing preparation can be effectively controlled. Meanwhile, the construction method is simple and efficient, the detection time can be shortened, the use of a solvent is reduced, the detection cost is further greatly reduced, and the method has high specificity and selectivity on analysis of unknown and known components and is beneficial to actual production and application.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, and more specifically, to the fingerprint spectrum and content determination method of a heart-nourishing and calming preparation for treating insomnia. Background Technology

[0002] The Heart-Nourishing and Mind-Calming Preparation is made from seven Chinese medicinal herbs—Acanthopanax senticosus, Platycladus orientalis seed, Ziziphus jujuba seed, Ginkgo biloba leaf, Longan pulp, Polygala tenuifolia, and mother-of-pearl—through extraction and purification. It has the effects of nourishing the heart, replenishing blood, and calming the mind. It is used for insomnia caused by deficiency of both heart and spleen, with symptoms such as difficulty falling asleep, decreased sleep quality and reduced sleep time, headache, dizziness, palpitations, forgetfulness, and excessive dreaming; and for neurasthenia with the above symptoms.

[0003] The existing quality standards for Yixin Anshen preparations include thin-layer chromatography identification of Acanthopanax senticosus, Ginkgo biloba leaf, Ziziphus jujuba seed, Longan pulp and Polygala tenuifolia, as well as content determination methods for Acanthopanax senticosus and Polygala tenuifolia. However, the thin-layer chromatography identification method cannot accurately control the quantitative quality of the product, and the content determination of single indicators such as syringin and 3,6'-disinoyl sucrose cannot comprehensively control and evaluate the product quality.

[0004] The applicant has developed a fingerprinting method using high-performance liquid chromatography (HPLC) for quality control of all five ingredients in a traditional Chinese medicine formula, enabling more comprehensive quality control. The chromatographic method developed by the applicant can rapidly and accurately separate seven active ingredients in the product: protocatechuic acid, syringin, chlorogenic acid, Siberian polygalactosyl sucrose A5, magnoflorine, spinosin, and 3,6'-disinoyl sucrose. The separation effect is excellent. Simultaneously, it can accurately quantify the active ingredients syringin, spinosin, and 3,6'-disinoyl sucrose contained in Acanthopanax senticosus, Ziziphus jujuba var. spinosa, and Polygala tenuifolia. The linear relationships of each ingredient are good, and the recovery rate is also good, meeting the technical requirements of "one test, multiple evaluations".

[0005] In view of this, the present invention provides a fingerprint spectrum and a method for determining the content of a heart-nourishing and calming preparation for treating insomnia. Summary of the Invention

[0006] The purpose of this invention is to provide a fingerprint spectrum and content determination method for a heart-nourishing and calming preparation for treating insomnia.

[0007] In order to achieve the above-mentioned objectives of the present invention, the following technical solution is adopted: In a first aspect, the present invention provides a method for constructing a fingerprint spectrum of a cardiac and sedative preparation for treating insomnia, the method comprising the following steps: The test solution containing the aforementioned euthanasia and sedation preparation was detected by high-performance liquid chromatography (HPLC), and the detection results were used to generate a fingerprint chromatogram. The HPLC detection used acetonitrile as mobile phase A and 0.1% aqueous phosphoric acid solution as mobile phase B. Gradient elution was performed using mobile phases A and B under the following conditions:

[0008] The percentages in the table represent the percentage of each mobile phase in the total volume of the mobile phase.

[0009] Furthermore, in this high-performance liquid chromatography detection, the chromatographic column is an octadecylsilane-bonded silica column, YMC-Pack ODS-AQ; the preferred specifications of the octadecylsilane-bonded silica column are 4.6 mm × 250 mm and 5 μm. Furthermore, in this high-performance liquid chromatography detection, the column temperature is 25–35°C; Furthermore, the detection wavelength of this high-performance liquid chromatography is 265–335 nm; Furthermore, the injection volume of this high-performance liquid chromatography is 5–20 μL, preferably 10 μL.

[0010] Furthermore, the flow rate of the mobile phase is 0.9–1.1 ml / min, preferably 1.0 ml / min.

[0011] Furthermore, it also includes preparing a reference solution and performing high-performance liquid chromatography on the reference solution to determine the peak assignment in the test solution based on the retention time of the reference solution; the reference solution includes protocatechuic acid, syringin, chlorogenic acid, Siberian polygalactosyl sucrose A5, magnoflorine, spinosin, and 3,6'-disinoyl sucrose.

[0012] Furthermore, the construction method includes the following steps: (1) Preparation of test solution: Dissolve the heart-nourishing and calming preparation in 50% methanol; (2) High performance liquid chromatography (HPLC) detection was performed using a YMC-Pack ODS-AQ column; the mobile phase was acetonitrile-0.1% phosphoric acid aqueous solution, the column temperature was 30℃, the flow rate was 1.0 mL / min, and the detection wavelengths were 265 nm, 320 nm and 335 nm. The gradient elution procedure is as follows:

[0013] The percentages in the table represent the percentage of each mobile phase in the total volume of the mobile phase.

[0014] Furthermore, step (1) also includes sonicating for 30 minutes (power 500W, frequency 40kHz), cooling, shaking, and filtering.

[0015] Furthermore, step (1) also includes the YMC-Pack ODS-AQ column having dimensions of 4.6 mm × 250 mm and 5 μm.

[0016] Secondly, the present invention provides a fingerprint spectrum of a heart-nourishing and tranquilizing preparation for treating insomnia. This fingerprint spectrum includes seven common characteristic peaks, with peak number 8 serving as a control peak. The relative retention times of the seven common characteristic peaks are as follows: The characteristic peak of No. 1 is protocatechuic acid, with a relative retention time of 0.414±10%; The characteristic peak of peak 5 is syringin, with a relative retention time of 0.764±10%. The characteristic peak of 6 is chlorogenic acid, with a relative retention time of 0.801±10%; The characteristic peak at point 7 is Siberian polygalactose A5, with a relative retention time of 0.817 ± 10%. The characteristic peak of peak 8 is magnoflorine, with a relative retention time of 1.000±10%. The characteristic peak at point 12 is a spinosine with a relative retention time of 1.465 ± 10%. The characteristic peak of nucleotide 16 is 3,6-disinyl sucrose, with a relative retention time of 1.807±10%.

[0017] Furthermore, the fingerprint spectrum of this heart-nourishing and calming preparation also includes 11 common characteristic peaks. With peak number 8 as the control peak, the relative retention times of the 11 common characteristic peaks are as follows: The relative retention time of the common characteristic peak No. 2 is 0.536 ± 10%; The relative retention time of the common characteristic peak No. 3 is 0.617 ± 10%; The relative retention time of the common characteristic peak No. 4 is 0.651 ± 10%; The relative retention time of the common characteristic peak at peak 9 is 1.223 ± 10%. The relative retention time of the common characteristic peak No. 10 is 1.380±10%; The relative retention time of the common characteristic peak No. 11 is 1.424 ± 10%; The relative retention time of the common characteristic peak 13 is 1.503 ± 10%; The relative retention time of the common characteristic peak 14 is 1.705 ± 10%; The relative retention time of the common characteristic peak No. 15 is 1.751 ± 10%; The relative retention time of the common characteristic peak No. 17 is 1.954 ± 10%; The relative retention time of the common characteristic peak at peak 18 is 2.070 ± 10%.

[0018] Thirdly, this invention provides a fingerprint spectrum of a heart-nourishing and tranquilizing preparation for treating insomnia, which is used as a standard fingerprint spectrum in the detection and quality control of the heart-nourishing and tranquilizing preparation.

[0019] Compared with the prior art, the beneficial effects of the present invention include at least the following: This invention provides the establishment and application of a fingerprint spectrum detection method for a cardiac and sedative preparation for treating insomnia. This fingerprint spectrum can simultaneously detect multiple chemical components in the cardiac and sedative preparation with high separation, systematically and stably reflecting the product quality and thus effectively controlling its quality. Furthermore, the construction method is simple and efficient, shortening detection time and reducing solvent usage, thereby significantly lowering detection costs. It also exhibits high specificity and selectivity for the analysis of both unknown and known components, making it more suitable for practical production applications. Attached Figure Description

[0020] Figure 1 Graph of Protocatechuic Acid Reference Standard Figure 2 Spectrum of Syringin Reference Standard Figure 3 Chlorogenic acid reference standard chromatogram Figure 4 Graph of Siberian Polygalactosyl Glucose A5 Reference Standard Figure 5 Spectrum of Magnoliaine Reference Standard Figure 6 Spinosin reference standard chromatogram Figure 7 3,6'-Disinosylsucrose reference standard chromatogram Figure 8 : Mixed reference standard chromatogram Figure 9 Spectrum of the test sample Figure 10 Spectrum of Acanthopanax senticosus extract Figure 11 Graph of Ziziphus jujuba seed extract Figure 12 Spectrum of Arborvitae Seed Extract Figure 13 Ginkgo biloba extract spectrum Figure 14 Spectrum of Polygala tenuifolia extract Figure 15 Graph of Longan Pulp Extract Figure 16 : Mixed reference standard spectrum—265nm Figure 17 Spectrum of the test sample—265nm Figure 18 Mixed reference standard chromatogram—320nm Figure 19 Spectrum of the test sample—320nm Figure 20 : Mixed reference standard spectrum—335nm Figure 21 Spectrum of the test sample—335nm Figure 22 Fingerprint spectrum of Yixin Anshen tablets Figure 23 Fingerprint profiles of 20 batches of test samples Detailed Implementation The embodiments of the present invention will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of the invention. Unless otherwise specified in the examples, conventional conditions or conditions recommended by the manufacturer are followed. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0021] The features and performance of the present invention will be further described in detail below with reference to embodiments: The method for constructing the fingerprint spectrum of a heart-nourishing and calming preparation includes the following steps: (1) Types of mobile phase: Acetonitrile is used as mobile phase A, and 0.1% phosphoric acid solution is used as mobile phase B; (2) The chromatographic column was: YMC-Pack ODS-AQ, 4.6 mm × 250 mm, 5 µm. (3) Flow rate: 1.0 ml / min (4) Column temperature: 25℃~35℃ (5) Detection wavelengths: fingerprint spectrum full spectrum 265nm; Acanthopanax senticosus content determination 265nm; Polygala tenuifolia content determination 320nm; Ziziphus jujuba seed content determination 335nm.

[0022] (6) The gradient elution ratio is:

[0023] (7) Preparation method of reference solution: Take an appropriate amount of magnoflorine reference standard, weigh it accurately, add 50% methanol to prepare a solution containing 0.1 mg per 1 ml, shake well, and it is ready.

[0024] (8) Preparation of mixed reference solutions: Take appropriate amounts of protocatechuic acid, syringin, chlorogenic acid, Siberian polygalactosyl sugar A5, magnoflorine, spinosin and 3,6'-disinoyl sucrose reference standards, accurately weigh them, add 50% methanol to dissolve them, and prepare solutions containing 5 μg of protocatechuic acid, syringin and Siberian polygalactosyl sugar A5 per 1 ml; 20 μg of chlorogenic acid and spinosin; 80 μg of magnoflorine; and 40 μg of 3,6'-disinoyl sucrose per 1 ml.

[0025] (9) Take an appropriate amount of this product, grind it into a fine powder, take about 1.0g, place it in a stoppered conical flask, add 25ml of 50% methanol, sonicate for 30 minutes (power 500W, frequency 40kHz), cool, shake well, filter, and take the filtrate to obtain the product.

[0026] (10) Determination method: Accurately pipette 10 μl of each of the reference solution, mixed reference solution and test solution into the liquid chromatograph, determine and record the chromatogram after 90 minutes.

[0027] The fingerprint chromatogram of the test sample should show 18 chromatographic peaks corresponding to the reference fingerprint chromatogram, of which peak 8 corresponds to the retention time of the reference peak; the similarity between the fingerprint chromatogram of the test sample and the reference fingerprint chromatogram calculated according to the similarity evaluation system of traditional Chinese medicine fingerprint chromatogram should not be less than 0.90.

[0028] The contents of syringin, spinosin, and 3,6'-disinoyl sucrose in the test sample were calculated using the external standard method.

[0029] 1. Establishment of fingerprinting method for heart-nourishing and calming preparations The specific steps of the fingerprint pattern construction method for the heart-nourishing and calming preparation in this embodiment are as follows: (1) Preparation of reference solutions for peak identification: Take appropriate amounts of protocatechuic acid, syringin, chlorogenic acid, Siberian polygalactosyl sugar A5, magnoflorine, spinosin and 3,6'-disinoyl sucrose reference standards, accurately weigh them, add 50% methanol to dissolve them, and prepare 0.1 mg of each of the following reference standards per 1 ml: protocatechuic acid, syringin, chlorogenic acid, Siberian polygalactosyl sugar A5, magnoflorine, spinosin and 3,6'-disinoyl sucrose.

[0030] (2) Preparation of mixed reference solution: Accurately pipette an appropriate amount of the above 6 reference stock solutions, add 50% methanol to prepare a solution containing 5 μg of protocatechuic acid, syringin, and Siberian polygalactosyl 5 per ml; 20 μg of chlorogenic acid and spinosin; 80 μg of magnoflorine; and 40 μg of 3,6'-disinoyl sucrose.

[0031] (3) Preparation of test solution: Take an appropriate amount of this product, grind it into a fine powder, take about 1.0g, place it in a stoppered conical flask, add 25ml of 50% methanol, sonicate for 30 minutes (power 500W, frequency 40kHz), cool, shake well, filter, and take the filtrate to obtain the test solution.

[0032] (4) Chromatographic conditions and system suitability test: Thermo Vanquish high performance liquid chromatograph, YMC-Pack ODS-AQ column; acetonitrile as mobile phase A, 0.1% phosphoric acid solution as mobile phase B, flow rate 1.0 ml / min; column temperature 30℃; detection wavelengths 265 nm, 320 nm and 335 nm. The theoretical plate number calculated based on the 3,6'-disinyl sucrose peak should not be less than 5000.

[0033] The high-performance liquid chromatography method employs gradient elution, and the elution program is as follows: 0–10 min, mobile phase A 5%, mobile phase B 95%; 10–40 min, mobile phase A 5%–15%, mobile phase B 95%–85%; 40–50 min, mobile phase A 15%, mobile phase B 85%; 50–80 min, mobile phase A 15%–25%, mobile phase B 85%–75%; 80–90 min, mobile phase A 25%, mobile phase B 75%; 90–95 min, mobile phase A 25%–20%, mobile phase B 75%–80%; 95–96 min, mobile phase A 20%–50%, mobile phase B 80%–50%; 96–100 min, mobile phase A 50%, mobile phase B 50%; 100–101 min, mobile phase A 50%–5%, mobile phase B 50%–95%; 101–105 min, mobile phase A 5%, mobile phase B 95%; (5) The detection wavelengths of the high performance liquid chromatography method are three wavelengths: 265nm, 320nm and 335nm.

[0034] Fingerprint spectrum was detected at 265 nm wavelength; syringin content was detected at 265 nm wavelength; 3,6'-disinoyl sucrose content was detected at 320 nm wavelength; and spinosin content was detected at 335 nm wavelength.

[0035] (6) Accurately pipette 10 µl of the reference solution and the test solution into the liquid chromatograph, measure and record the chromatogram after 90 minutes.

[0036] (7) See attached chromatogram. Figure 1-9 . (8) Relative retention time (min) of each common peak

[0037] (9) Similarity between the fingerprint chromatograms of each batch of test samples and the control.

[0038]

[0039] (10) Figure 9 The chromatogram of the test sample was obtained by comparing it with... Figures 1 to 8 The chromatogram of the test sample was compared with the reference standard. The chromatographic peaks in the chromatogram of the test sample were as follows: Peak 1: protocatechuic acid, Peak 5: syringin, Peak 6: chlorogenic acid, Peak 7: Siberian polygalactosyl sugar A5, Peak 8: magnoflorine, Peak 12: spinosin, Peak 16: 3,6-disinoyl sucrose, Peaks 2-4, 9-11, 13-15, and 17-18: unknown peaks. The chromatogram showed good separation of each peak without tailing or crossover. It can simultaneously detect the fingerprint spectrum and the content of the effective components of Acanthopanax senticosus, Ziziphus jujuba var. spinosa, and Polygala tenuifolia, and can be used for comprehensive quality evaluation of the heart-nourishing and calming preparation.

[0040] 2. Attribution of chromatographic peaks in the fingerprint chromatogram of the heart-nourishing and calming preparation. (1) Experimental method: Take the single herb extracts of cypress seed, longan pulp, polygala root, ginkgo leaf, jujube seed, and eleutherococcus senticosus, respectively, and process them according to the test sample solution preparation method in Example 1 to obtain the corresponding single herb test sample solutions.

[0041] (2) Following the fingerprint analysis method in Example 1, the solutions of each single herb were injected into a high-performance liquid chromatograph for analysis to determine peak assignment. The chromatograms are attached. Figure 9-15 . (3) Research Results Peak attribution study results table

[0042] (4) From the appendix Figure 9-15 It can be seen that peak 2 (unknown peak), peak 14 (unknown peak), and peak 15 (unknown peak) are characteristic peaks specific to Ginkgo biloba leaves; peak 12 (spinosin) is a characteristic peak of Ziziphus jujuba seed; peak 3 (unknown peak), peak 7 (Siberian polygalactosyl sugar A5), peak 10 (unknown peak), peak 16 (3,6'-disinyl sucrose), peak 17 (unknown peak), and peak 18 (unknown peak) are characteristic peaks specific to Polygala tenuifolia; peak 5 (syringin) and peak 6 (chlorogenic acid) are characteristic peaks specific to Acanthopanax senticosus.

[0043] Peak 1 (protocatechuic acid) is a common peak of Arborvitae seed, Ginkgo leaf and Acanthopanax senticosus; Peak 4 (unknown peak) and Peak 11 (unknown peak) are common peaks of Ziziphus jujuba seed, Ginkgo leaf and Polygala tenuifolia; Peak 8 (magnolia alkaloid) is a common peak of Ziziphus jujuba seed and Polygala tenuifolia; Peak 9 (unknown peak) and Peak 13 (unknown peak) are common peaks of Ginkgo leaf and Polygala tenuifolia; 18 common peaks cover all 5 herbs in the prescription. This method can achieve quality control of the 5 herbs in the prescription: Acanthopanax senticosus, Ziziphus jujuba seed, Ginkgo leaf, Polygala tenuifolia and Arborvitae seed.

[0044] 3. Linearity test of the heart-nourishing and calming preparation (1) Test method: Weigh approximately 10.53 mg of syringin reference standard (batch number: 111574-202106, content: 94.3%) accurately, place it in a 100 ml volumetric flask, add 50% methanol solution and dilute to the mark, shake well, and use as the syringin reference standard stock solution. Accurately pipette the syringin reference standard stock solution and dilute it with 50% methanol solution to prepare solutions containing 1 μg, 3 μg, 5 μg, 7 μg and 9 μg per ml, respectively, as syringin linear reference standard solutions.

[0045] Accurately weigh approximately 15.67 mg of spinosine reference standard (batch number: 111869-202406, purity: 96.6%), place it in a 50 ml volumetric flask, add 50% methanol solution and dilute to the mark, shake well, and use as the spinosine reference standard stock solution. Accurately pipette the spinosine reference standard stock solution and dilute with 50% methanol solution to prepare solutions containing 3 μg, 9 μg, 15 μg, 21 μg, and 27 μg per ml, respectively, to prepare as linear spinosine reference solutions.

[0046] Accurately weigh approximately 20.06 mg of 3,6'-disinoyl sucrose reference standard (batch number: 111848-202307, purity: 96.7%), place it in a 50 ml volumetric flask, add 50% methanol solution and dilute to the mark, shake well, and prepare the paeoniflorin reference standard stock solution. Accurately pipette the reference standard stock solution and dilute with 50% methanol solution to prepare solutions containing 20 μg, 40 μg, 60 μg, 80 μg, and 100 μg per ml, respectively, to prepare the 3,6'-disinoyl sucrose linear reference standard solutions.

[0047] (2) The above series of concentration reference solutions were detected under the chromatographic conditions of Example 1. A standard curve was plotted with the reference concentration as the abscissa and the peak area as the ordinate, and the linear regression equation and correlation coefficient were calculated.

[0048] (3) Linear correlation of syringin

[0049]

[0050] (4) Linear correlation of Spinosin

[0051]

[0052] (5) Linear correlation of 3,6'-disinyl sucrose

[0053]

[0054] (6) The above data show that this method has a good linear correlation in detecting the content of syringin, spinosin and 3,6'-disinoyl sucrose.

[0055] 4. Accuracy test of the heart-nourishing and calming preparation (1) Test method: In this example, the recovery rate test was carried out using reference standards. Syringin, spinosin and 3,6'-disinoyl sucrose reference standards were added to the test sample with known contents of syringin, spinosin and 3,6'-disinoyl sucrose for determination, and the recovery rate of each added reference standard was calculated.

[0056] (2) Preparation of test solution: First, prepare a stock solution containing syringin, spinosin and 3,6'-disinoyl sucrose reference standard. Add the stock solution of reference standard and the test solution at ratios of 1:0.5, 1:1 and 1:1.5 to prepare test solutions of low, medium and high concentrations.

[0057] (3) Preparation of reference solution: Accurately weigh appropriate amounts of syringin, spinosin and 3,6'-disinoyl sucrose reference standards, add 50% methanol to prepare a mixed solution containing 65 μg of syringin, 110 μg of spinosin and 360 μg of 3,6'-disinoyl sucrose per ml.

[0058] (4) Accurately pipette 10 µl of the reference solution and the test solution into the liquid chromatograph and determine them under the chromatographic conditions of Example 1. Calculate the content of the test sample using the external standard method and calculate the recovery rate. The results are shown in Table 1-3. (5) Formula for calculating recovery rate

[0059] (6) Table 1 Recovery rate of syringin

[0060] (7) Table 2 Spinosin recovery rate

[0061] (8) Table 33,6'-Disinylsucrose recovery rate

[0062] (9) According to the above recovery data, the recovery rates of syringin, spinosin and 3,6'-disinoyl sucrose by this method are all between 90% and 108%, which meets the requirements of the Pharmacopoeia's "Guiding Principles for Validation of Analytical Methods".

[0063] All aspects not described in detail in this invention are common knowledge that can be readily understood by those skilled in the art. Although the invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which are obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of this invention fall within the scope of protection claimed by this invention.

Claims

1. A method for constructing the fingerprint spectrum of a heart-nourishing and calming preparation for treating insomnia, characterized in that, The construction method includes the following steps: The test solution containing the aforementioned euthanasia and sedation preparation was detected by high-performance liquid chromatography (HPLC), and the detection results were used to generate a fingerprint chromatogram. The HPLC detection used acetonitrile as mobile phase A and 0.1% aqueous phosphoric acid solution as mobile phase B. Gradient elution was performed using mobile phases A and B under the following conditions: The percentages in the table represent the percentage of each mobile phase in the total volume of the mobile phase.

2. The construction method according to claim 1, characterized in that, In the high-performance liquid chromatography (HPLC) detection, the chromatographic column is an octadecylsilane-bonded silica column, YMC-Pack ODS-AQ; the preferred specifications of the octadecylsilane-bonded silica column are 4.6 mm × 250 mm and 5 μm; the column temperature is 25–35 °C; the detection wavelength is 265–335 nm; the flow rate of the mobile phase is 0.9–1.1 ml / min; and the injection volume is 5–20 μL.

3. The construction method according to claim 1 or 2, characterized in that, In the high-performance liquid chromatography (HPLC) detection, the column temperature is 30℃; the flow rate of the mobile phase is 1.0 ml / min; and the injection volume of the HPLC is 10 μL.

4. The construction method according to any one of claims 1-3, characterized in that, It also includes preparing a reference solution and performing high-performance liquid chromatography (HPLC) on the reference solution to determine the peak assignment in the test solution based on the retention time of the reference solution; the reference solution includes protocatechuic acid, syringin, chlorogenic acid, Siberian polygalactosyl sucrose A5, magnoflorine, spinosin, and 3,6'-disinoyl sucrose.

5. The construction method according to any one of claims 1-4, characterized in that, The construction method includes the following steps: (1) Preparation of test solution: Dissolve the heart-nourishing and calming preparation in 50% methanol; (2) High performance liquid chromatography (HPLC) detection was performed using a YMC-Pack ODS-AQ column; the mobile phase was acetonitrile-0.1% phosphoric acid aqueous solution, the column temperature was 30℃, the flow rate was 1.0 mL / min, and the detection wavelengths were 265 nm, 320 nm and 335 nm. The gradient elution procedure is as follows: The percentages in the table represent the percentage of each mobile phase in the total volume of the mobile phase.

6. The construction method according to claim 5, characterized in that, Step (1) also includes sonicating for 30 minutes (power 500W, frequency 40kHz), cooling, shaking, and filtering.

7. The construction method according to claim 5, characterized in that, Step (1) also includes the YMC-PackODS-AQ chromatographic column having a specification of 4.6mm×250mm and 5μm.

8. A fingerprint spectrum of a heart-nourishing and calming preparation for treating insomnia, characterized in that, The fingerprint spectrum includes 7 common characteristic peaks, with peak number 8 as the control peak. The relative retention times of the 7 common characteristic peaks are as follows: The characteristic peak of No. 1 is protocatechuic acid, with a relative retention time of 0.414±10%; The characteristic peak of peak 5 is syringin, with a relative retention time of 0.764±10%. The characteristic peak of 6 is chlorogenic acid, with a relative retention time of 0.801±10%; The characteristic peak at point 7 is Siberian polygalactose A5, with a relative retention time of 0.817 ± 10%. The characteristic peak of peak 8 is magnoflorine, with a relative retention time of 1.000±10%. The characteristic peak at point 12 is a spinosine with a relative retention time of 1.465 ± 10%. The characteristic peak of nucleotide 16 is 3,6-disinyl sucrose, with a relative retention time of 1.807±10%.

9. The fingerprint spectrum according to claim 8, characterized in that, The fingerprint spectrum of the heart-nourishing and calming preparation also includes 11 common characteristic peaks, with peak number 8 as the control peak. The relative retention times of the 11 common characteristic peaks are as follows: The relative retention time of the common characteristic peak No. 2 is 0.536 ± 10%; The relative retention time of the common characteristic peak No. 3 is 0.617 ± 10%; The relative retention time of the common characteristic peak No. 4 is 0.651 ± 10%; The relative retention time of the common characteristic peak at peak 9 is 1.223 ± 10%. The relative retention time of the common characteristic peak No. 10 is 1.380±10%; The relative retention time of the common characteristic peak No. 11 is 1.424 ± 10%; The relative retention time of the common characteristic peak 13 is 1.503 ± 10%; The relative retention time of the common characteristic peak 14 is 1.705 ± 10%; The relative retention time of the common characteristic peak No. 15 is 1.751 ± 10%; The relative retention time of the common characteristic peak No. 17 is 1.954 ± 10%; The relative retention time of the common characteristic peak at peak 18 is 2.070 ± 10%.

10. The application of a fingerprint spectrum of the heart-nourishing and calming preparation according to claim 8 or 9 as a standard fingerprint spectrum in the detection and quality control of the heart-nourishing and calming preparation.

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