Method for constructing shengpi bagged tea fingerprint

Abstract

The application discloses a kind of construction methods of Shengmai teabag fingerprint, comprising the following steps: reference solution preparation, test sample solution preparation, precision pipetting reference solution and test sample solution are injected into high performance liquid chromatograph, and chromatogram is recorded.The Shengmai teabag fingerprint constructed by the application calibrates 15 common characteristic peaks, and identifies the chemical composition of 6 characteristic peaks, can simultaneously reflect the overall situation of multiple active ingredients in product.The methodological verification result shows that the method has strong specificity, good precision and good stability, the obtained fingerprint information is complete, has good separation degree and smooth baseline, can accurately reflect the quality characteristics of Shengmai teabag, and can be used for the overall quality evaluation of Shengmai teabag, effectively guarantee the consistency and stability of product quality.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine analysis technology, and in particular relates to a method for constructing a fingerprint spectrum of Shengmai tea bags. Background Technology

[0002] Shengmai tea bags are a traditional Chinese medicine compound preparation composed of ginseng, ophiopogon japonicus, and schisandra chinensis. They possess the effects of invigorating qi, nourishing yin, and promoting body fluid production, and are used for qi and yin deficiency, palpitations, shortness of breath, and spontaneous sweating. Shengmai tea bags comply with the National Drug Standard YBZ03212008 issued by the State Drug Administration. However, the current standard lacks a quantitative-quality transfer study of multi-component information, which is detrimental to product quality control. Therefore, this study aims to develop a fingerprinting method for Shengmai tea bags. This method can more comprehensively control product quality, accurately evaluate the overall quality consistency of Shengmai tea bags, and better ensure the clinical safety and stable efficacy of this drug. Summary of the Invention

[0003] This invention provides a method for constructing a fingerprint spectrum of Shengmai tea bags. Compared with the current quality standards for single-component detection, this method can simultaneously cover information on multiple characteristic chemical components in Shengmai tea bags, enabling control over the overall quality of the product. It fills the gap in the existing quality standards for Shengmai tea bags regarding the overall quality control of multiple components and provides reliable technical support for improving the quality standardization of this product.

[0004] This invention provides a method for constructing a fingerprint spectrum of Shengmai tea bags, comprising the following steps:

[0005] (1) Preparation of reference solution: Weigh 5-hydroxymethylfurfural, 4-coumaric acid, schisandrol A, schisandrol B, schisandrin A, and schisandrin B accurately, dissolve them in methanol solution to obtain reference solution;

[0006] (2) Preparation of test solution: Take an appropriate amount of Shengmai tea bag, grind it into a fine powder, add methanol solution, weigh it, sonicate it, cool it, make up the weight loss with methanol solution, shake it well, filter it, and take the filtrate to obtain the test solution.

[0007] (3) Determination: Inject the reference solution and the test solution into the high performance liquid chromatograph, determine and record the chromatogram;

[0008] The chromatographic conditions of the high-performance liquid chromatograph are as follows:

[0009] Chromatographic column: C18 column, 250mm × 4.6mm × 5μm;

[0010] Mobile phase A: Acetonitrile;

[0011] Mobile phase B: 0.2% glacial acetic acid solution;

[0012] The mobile phase flow rate was 0.7~1.1 ml / min;

[0013] Column temperature: 28~32℃;

[0014] Detection wavelength: 270~290nm;

[0015] The detector is an ultraviolet detector;

[0016] The gradient elution procedure is as follows:

[0017] At 0 minutes, mobile phase A is 5% and mobile phase B is 95%.

[0018] At 10 minutes, mobile phase A was 5% and mobile phase B was 95%.

[0019] At 45 minutes, mobile phase A was 19% and mobile phase B was 81%.

[0020] At 60 minutes, mobile phase A was 45% and mobile phase B was 55%.

[0021] At 65 minutes, mobile phase A was 50% and mobile phase B was 50%.

[0022] At 79 minutes, mobile phase A was 50% and mobile phase B was 50%.

[0023] At 85 minutes, mobile phase A was 70% and mobile phase B was 30%.

[0024] At 105 minutes, mobile phase A was 72% and mobile phase B was 28%.

[0025] At 110 minutes, mobile phase A was 5% and mobile phase B was 95%.

[0026] At 120 minutes, mobile phase A was 5% and mobile phase B was 95%.

[0027] The methanol solution is a 50% aqueous methanol solution.

[0028] The fingerprint spectrum of the Shengmai tea bag includes 15 marker peaks, of which peak 1 is 5-hydroxymethylfurfural, peak 5 is 4-coumaric acid, peak 8 is schisandrol A, peak 11 is schisandrol B, peak 14 is schisandrin A, and peak 15 is schisandrin B.

[0029] This invention establishes a fingerprint spectrum for Shengmai tea bags, identifying 15 common characteristic peaks and pinpointing the chemical components of 6 of these peaks. This simultaneously reflects the overall composition of multiple active ingredients in the product. Methodological validation results demonstrate that this method is highly specific, precise, and stable. The obtained fingerprint spectrum information is complete, with good separation and a stable baseline, accurately reflecting the quality characteristics of Shengmai tea bags. It can be used for the overall quality evaluation of Shengmai tea bags, effectively ensuring the consistency and stability of product quality. Attached Figure Description

[0030] Figure 1 Wavelength analysis of chromatograms;

[0031] Figure 2 The chromatogram was examined based on the detection time of the test sample.

[0032] Figure 3 Peak localization and attribution chromatogram of the test sample;

[0033] Figure 4 Chromatogram of peak labeling of the test sample;

[0034] Figure 5 Flow rate analysis of chromatograms;

[0035] Figure 6 Chromatograms of the extraction solvent for the test sample;

[0036] Figure 7 Comparison of chromatograms for system suitability and instrument precision similarity;

[0037] Figure 8 Specificity comparison chromatogram;

[0038] Figure 9 Control fingerprints generated from repeatable experiments;

[0039] Figure 10 Chromatograms for repeatability similarity comparison;

[0040] Figure 11 Intermediate precision similarity comparison chromatogram;

[0041] Figure 12 Chromatograms comparing the similarity between the control fingerprint chromatogram and the intermediate precision chromatogram generated from repeatability experiments;

[0042] Figure 13 Chromatograms comparing similarity at different wavelengths;

[0043] Figure 14 Chromatograms showing similarity at different flow rates;

[0044] Figure 15 Chromatograms showing similarity at different column temperatures;

[0045] Figure 16 Stability similarity comparison chromatograms. Detailed Implementation

[0046] To enable those skilled in the art to more clearly understand the technical solutions described in this invention, the following embodiments are provided for illustration. It should be noted that the following embodiments do not constitute a limitation on the scope of protection claimed by this invention.

[0047] Unless otherwise specified, the raw materials, reagents or devices used in the following examples are available from conventional commercial sources or can be obtained by existing known methods.

[0048] Example 1: Investigation of Chromatographic Conditions

[0049] (1) Wavelength investigation

[0050] Preparation of reference solution: Take appropriate amounts of schisandrol A, schisandrin A, schisandrin B, schisandrol B, 5-hydroxymethylfurfural and 4-coumaric acid reference standards, accurately weigh them, add 50% methanol to prepare a solution containing 100 μg per ml.

[0051] Preparation of the test solution: Take an appropriate amount of this product, grind it into a fine powder, take about 2.5g, weigh it accurately, place it in a stoppered conical flask, accurately add 20ml of 50% methanol, weigh it, sonicate for 30 minutes, take it out, let it cool, weigh it again, make up the lost weight with 50% methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0052] Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the high performance liquid chromatograph and determine the result.

[0053] The chromatographic conditions were as follows: a C18 column, 250 mm × 4.6 mm × 5 μm, with acetonitrile as mobile phase A and 0.2% glacial acetic acid solution as mobile phase B, and gradient elution as specified in the table below; the flow rate was 1 ml / min; the column temperature was 30 ℃; and detection was performed using a UV detector at wavelengths of 240 nm, 260 nm, 270 nm, 280 nm, 290 nm, and 295 nm.

[0054]

[0055] Chromatogram (see) Figure 1 The results showed that when the detection wavelength of the test sample was below 270 nm (excluding 270 nm), the peak baseline was uneven and the peak response was small when the retention time was 20-50 min; when the detection wavelength of the test sample was above 290 nm (excluding 290 nm), the peak response of schisandrin was small when the retention time was about 92 min. The peak distribution was more uniform and the response was better when the wavelength was between 270 nm and 290 nm.

[0056] (2) Examination of detection duration

[0057] The preparation of the reference solution, the preparation of the test solution, and the high-performance liquid chromatography conditions were the same as those for the wavelength investigation test, except for the gradient elution program, which is shown in Table 2.

[0058]

[0059] Chromatogram (see) Figure 2 .from Figure 2 As can be seen from the data, the main components contained in Shengmai tea bags all reached their peaks within 120 minutes, so a detection time of 120 minutes is sufficient to meet the detection requirements.

[0060] (3) Chromatographic peak assignment and identification

[0061] Preparation of Schisandra chinensis negative (Schisandra chinensis deficient) sample: Take about 100g of ginseng and about 200g of Ophiopogon japonicus that have been crushed into coarse powder, mix them evenly, make them into granules, and dry them to obtain the sample.

[0062] Preparation of ginseng-negative (ginseng-deficient) samples: Take about 100g of coarsely ground Ophiopogon japonicus and about 50g of Schisandra chinensis, mix them evenly, make them into granules, and dry them to obtain the sample.

[0063] Preparation of Ophiopogon japonicus negative (Ophiopogon japonicus deficient) sample: Take about 50g of ginseng and about 50g of Schisandra chinensis, which are crushed into coarse powder, mix them evenly, make them into granules, and dry them to obtain the sample.

[0064] Preparation of Schisandra chinensis negative (Schisandra chinensis absent) sample test solution: Take an appropriate amount of Schisandra chinensis negative sample, grind it into a fine powder, mix it well, take about 2.5g, accurately weigh it, place it in a stoppered conical flask, accurately add 20ml of 50% methanol, weigh it, sonicate for 30 minutes, take it out, let it cool, weigh it again, make up the lost weight with 50% methanol, shake it well, filter it, and take the filtrate to obtain the test solution.

[0065] Preparation of test solution for ginseng-negative (ginseng-deficient) sample: Take an appropriate amount of ginseng-negative sample, grind it into a fine powder, mix it well, take about 2.5g, weigh it accurately, place it in a stoppered conical flask, accurately add 20ml of 50% methanol, weigh it, sonicate it for 30 minutes, take it out, let it cool, weigh it again, make up the weight loss with 50% methanol, shake it well, filter it, and take the filtrate to obtain the solution.

[0066] Preparation of test solution for Ophiopogon japonicus negative (Ophiopogon japonicus deficient) sample: Take an appropriate amount of Ophiopogon japonicus negative sample, grind it finely, mix it well, take about 2.5g, weigh it accurately, put it in a stoppered conical flask, accurately add 20ml of 50% methanol, weigh it, sonicate for 30 minutes, take it out, let it cool, weigh it again, make up the lost weight with 50% methanol, shake it well, filter it, and take the filtrate to obtain the test solution.

[0067] Preparation of Schisandra chinensis test solution: Take about 2.5g of Schisandra chinensis powder, accurately weigh it, place it in a stoppered conical flask, accurately add 20ml of 50% methanol, weigh it, sonicate for 30 minutes, take it out, let it cool, weigh it again, make up the lost weight with 50% methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0068] Preparation of ginseng test solution: Take about 2.5g of ginseng powder, place it in a stoppered conical flask, accurately add 20ml of 50% methanol, weigh it, sonicate for 30 minutes, take it out, cool it, weigh it again, replenish the lost weight with 50% methanol, shake well, filter it, and take the filtrate to obtain the test solution.

[0069] Preparation of Ophiopogon japonicus test solution: Take about 2.5g of Ophiopogon japonicus powder, place it in a stoppered conical flask, accurately add 20ml of 50% methanol, weigh it, sonicate for 30 minutes, take it out, let it cool, weigh it again, make up the lost weight with 50% methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0070] The preparation of the reference solution, the test solution (sample), and the high-performance liquid chromatography conditions were consistent with the wavelength investigation test conditions.

[0071] Determination method: Accurately pipette 10 μl of each of the above-mentioned test solutions and reference solutions, inject them into the liquid chromatograph, and determine the result.

[0072] Chromatogram (see) Figure 3 and Figure 4 As shown in the figure, the peak assignments and peak locations in the chromatogram of Shengmai tea bags are shown in Table 3 below.

[0073]

[0074] (4) Flow velocity investigation

[0075] The preparation of the reference solution, the test solution, and the high-performance liquid chromatography (HPLC) conditions were consistent with the wavelength determination test conditions, with only the flow rate adjusted to 0.5 ml / min, 0.8 ml / min, 0.9 ml / min, 1 ml / min, 1.1 ml / min, and 1.2 ml / min. Chromatograms at different flow rates are shown below. Figure 5 The results showed that, under different flow rate conditions, the chromatographic peaks were best separated at a flow rate of 0.9 ml / min.

[0076] Example 2: Investigation of Extraction Solvent

[0077] Preparation of reference solution: Take appropriate amounts of schisandrol A, schisandrin A, schisandrin B, schisandrol B, 5-hydroxymethylfurfural and 4-coumaric acid reference standards, accurately weigh them, add 50% methanol to prepare a solution containing 100 μg per ml.

[0078] Preparation of the test solution: Take an appropriate amount of this product, grind it into a fine powder, take about 2.5g, weigh it accurately, place it in a stoppered conical flask, accurately add 20ml of 50% methanol, weigh it, sonicate for 30 minutes, take it out, let it cool, weigh it again, make up the lost weight with 50% methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0079] Preparation of test solution 1: Take an appropriate amount of this product, grind it into a fine powder, take about 2.5g, weigh it accurately, place it in a stoppered conical flask, accurately add 20ml of 30% methanol, weigh it, sonicate for 30 minutes, take it out, cool it, weigh it again, make up the lost weight with 30% methanol, shake it well, filter it, and take the filtrate to obtain the test solution.

[0080] Preparation of test solution 2: Take an appropriate amount of this product, grind it into a fine powder, take about 1.0 g, weigh it accurately, place it in a stoppered conical flask, accurately add 25 ml of 80% methanol, weigh it, sonicate for 30 minutes, take it out, cool it, weigh it again, make up the weight loss with 80% methanol, shake it well, filter it, and take the filtrate to obtain the test solution.

[0081] Assay: Accurately pipette 10 μl each of the reference solution and test solution 1 (i.e., the test solution prepared using the same method as test solution 1), and accurately pipette 30 μl of test solution 2 (i.e., the test solution prepared using the same method as test solution 2), inject them into the liquid chromatograph, and determine the result. The chromatographic conditions for the test are consistent with those for the wavelength investigation.

[0082] Chromatogram (see) Figure 6 ,from Figure 6 As can be seen from the data, when the extraction solvent is 50% methanol, the peak shape and peak response in the chromatogram are both good.

[0083] Example 3: Determination of reference standard concentration

[0084] Chromatographic conditions and system suitability test: The chromatographic column is a C18 column, 250 mm × 4.6 mm × 5 μm; acetonitrile is used as mobile phase A, and 0.2% glacial acetic acid solution is used as mobile phase B, and gradient elution is performed according to the specifications in the table below; the flow rate is 0.9 ml / min; the column temperature is 30 ℃, and the detection wavelength is 280 nm.

[0085]

[0086] Preparation of reference solution: Take appropriate amounts of schisandrol A, schisandrin A, schisandrin B, schisandrol B, 5-hydroxymethylfurfural and 4-coumaric acid reference standards, accurately weigh them, add 50% methanol to prepare a solution containing 100 μg per ml.

[0087] Preparation of the test solution: Take an appropriate amount of this product, grind it into a fine powder, take about 2.5g, weigh it accurately, place it in a stoppered conical flask, accurately add 20ml of 50% methanol, weigh it, sonicate for 30 minutes, take it out, let it cool, weigh it again, make up the lost weight with 50% methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0088] Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0089] The test results are shown in Table 5. Based on the test results in Table 5, in order to ensure that the peak areas of the test sample and the reference sample are comparable, the concentration of the reference sample was adjusted to contain 10 μg of 5-hydroxymethylfurfural, 5 μg of 4-coumaric acid, 330 μg of schisandrol A, 65 μg of schisandrol B, 80 μg of schisandrin A, and 45 μg of schisandrin B per 1 ml.

[0090]

[0091] Example 4 Methodological Validation

[0092] (1) System applicability and instrument precision

[0093] Blank solution: 50% methanol.

[0094] Test solution: Take an appropriate amount of this product, grind it into a fine powder, take about 2.5g, weigh it accurately, place it in a stoppered conical flask, accurately add 20ml of 50% methanol, weigh it, sonicate for 30 minutes, take it out, let it cool, weigh it again, make up the weight loss with 50% methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0095] Precisely inject 10 μl of blank solution into the liquid chromatograph and inject 10 μl of the test solution into the liquid chromatograph six times. Measure and record the chromatograms. Import the chromatograms of the test solution obtained from the six injections into the Chinese herbal chromatographic fingerprint similarity evaluation system to calculate the similarity.

[0096] Chromatogram (see) Figure 7The similarity comparison results are shown in Table 6. The results show that the similarity of the fingerprint spectra from six consecutive injections of the same test solution is above 0.992. Table 7 shows that for the same test solution, the retention time RSD of the schisandrol A peak was 0.0139%, the peak area RSD was 0.0389%, the resolution between the schisandrol A peak and adjacent peaks was ≥1.5, the tailing factor was between 0.8 and 1.8, and the theoretical plate number was not less than 2000, indicating good system applicability and instrument precision.

[0097]

[0098]

[0099] (2) Exclusivity

[0100] Blank solution: 50% methanol.

[0101] The preparation methods of the reference solution and the test solution are the same as those used in the wavelength investigation test of Example 1.

[0102] Accurately pipette 10 μl each of the blank solution, reference solution, and test solution, inject them into the liquid chromatograph, measure, and record the chromatograms. The chromatographic conditions are the same as those in the wavelength investigation test of Example 1.

[0103] Chromatogram (see) Figure 8 The results showed that the blank solution (50% methanol) chromatogram did not have a corresponding peak at the retention time corresponding to the reference standard; the test solution chromatogram showed a corresponding peak at the retention time corresponding to the reference standard, indicating good specificity.

[0104] (3) Precision

[0105] (3.1) Repeatability:

[0106] The blank solution, reference solution, and test solution were consistent with the specificity test. Six parallel tests were prepared for the test solution.

[0107] Accurately measure 10 μl each of the blank solution, reference solution, and test solution, inject them into the liquid chromatograph, determine the chromatograms, and record the chromatograms. The chromatographic conditions should be consistent with those of the specificity test. Import the chromatograms of the six test solutions into the Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (version 2012.130723), calculate the similarity, and generate the reference fingerprint. The repeatability similarity results are shown in Table 8.

[0108]

[0109] (3.2) Intermediate precision:

[0110] Verification experiments were conducted by different personnel at different times using different instruments, and the experimental content was the same as the verification process of repeatability experiments.

[0111] Repeatability and intermediate precision chromatograms are shown in Figures 9-12 The intermediate precision similarity results are shown in Table 9. The results show that the similarity between the fingerprint chromatograms of the test solution and the repeatability and intermediate precision chromatograms is above 0.992. The similarity between the fingerprint chromatograms of the six test solution generated in the intermediate precision experiment and the control fingerprint chromatogram generated in the repeatability experiment is above 0.906, indicating good precision.

[0112]

[0113]

[0114] (4) Durability

[0115] Blank solution: Take 50% methanol to obtain the blank solution.

[0116] Reference solution: Same as specificity test.

[0117] Test solution: Take an appropriate amount of this product, grind it into a fine powder, take about 2.5g, weigh it accurately, place it in a stoppered conical flask, accurately add 20ml of 50% methanol, weigh it, sonicate for 30 minutes, take it out, let it cool, weigh it again, make up the weight loss with 50% methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0118] Accurately pipette 10 μl each of the blank solution, reference solution, and test solution into the liquid chromatograph. Measure and record the chromatograms at a column temperature of 30℃±2℃, a wavelength of 280nm±5nm, and a flow rate of 0.9 ml / min±0.2 ml / min. Other chromatographic conditions should be consistent with the specificity test. Import the chromatograms of the test solution under the corresponding conditions into the Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (version 2012.130723) to calculate the similarity.

[0119] Fingerprint chromatograms of test solution chromatograms at different wavelengths are shown below. Figure 13 The similarity comparison results are shown in Table 11. By comparison, it can be seen that the similarity between the fingerprint spectra of 275nm and 280nm is 0.994, and the similarity between the fingerprint spectra of 285nm and 280nm is 0.995.

[0120]

[0121] Fingerprint chromatograms of test solutions at different flow rates are shown in [reference needed]. Figure 14The similarity comparison results are shown in Table 12. The results show that the fingerprint pattern similarity between 0.7 ml / min and 0.9 ml / min is 0.941, and the fingerprint pattern similarity between 1.1 ml / min and 0.9 ml / min is 0.936.

[0122]

[0123] Fingerprint chromatograms of test solution chromatograms at different column temperatures are shown below. Figure 15 The similarity comparison results are shown in Table 13. The results show that the similarity between the fingerprint spectra of 28℃ and 30℃ is 0.986, and the similarity between the fingerprint spectra of 32℃ and 30℃ is 0.960.

[0124]

[0125] (5) Solution stability:

[0126] Blank solution: Take 50% methanol to obtain the blank solution.

[0127] Reference solution: Same as specificity test.

[0128] Test solution: Take an appropriate amount of this product, grind it into a fine powder, take about 2.5g, weigh it accurately, place it in a stoppered conical flask, accurately add 20ml of 50% methanol, weigh it, sonicate for 30 minutes, take it out, let it cool, weigh it again, make up the weight loss with 50% methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0129] Accurately pipette 10 μl each of blank solution, reference solution, and test solution, and inject them into the liquid chromatograph. Analyze the test solution at different time points (at least up to 48 h) and record the chromatograms. Import the chromatograms of the test solution at different time points into the Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (version 2012.130723) and calculate the similarity.

[0130] The fingerprint chromatograms of the test solution after preparation and storage for different times are shown in the figure. Figure 16 The similarity results are shown in Table 14. The comparison of the results shows that the similarity of the 11 needles is above 0.970.

[0131]

[0132]

[0133] The results of the stability test of the test sample solution are shown in Table 15. As can be seen from Table 15, at different time points in the solution stability test, the retention time and peak area RSD of 5-hydroxymethylfurfural, 4-coumaric acid, schisandrol A, schisandrol B, schisandrin A, and schisandrin B in the test sample solution were all ≤5.0%, indicating that the test sample solution had good stability within 68 hours.

[0134] After systematic methodological validation, this fingerprint analysis method meets the requirements in terms of system applicability, instrument precision, specificity, and robustness. The similarity among all experimental results is no less than 0.90, and the RSD values ​​of the retention time and peak area of ​​schisandrol A in the chromatogram of the test sample are both ≤1.0% in the stability test. This method has demonstrated good methodological validation.

[0135] The above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A method for constructing a fingerprint spectrum of Shengmai tea bags, characterized in that, Includes the following steps: (1) Preparation of reference solution: Weigh 5-hydroxymethylfurfural, 4-coumaric acid, schisandrol A, schisandrol B, schisandrin A, and schisandrin B accurately, dissolve them in methanol solution to obtain reference solution; (2) Preparation of test solution: Take an appropriate amount of Shengmai tea bag, grind it into a fine powder, add methanol solution, weigh it, sonicate it, cool it, make up the weight loss with methanol solution, shake it well, filter it, and take the filtrate to obtain the test solution. (3) Determination: Inject the reference solution and the test solution into the high performance liquid chromatograph, determine and record the chromatogram; The chromatographic conditions of the high-performance liquid chromatograph are as follows: Chromatographic column: C18 column, 250mm × 4.6mm × 5μm; Mobile phase A: Acetonitrile; Mobile phase B: 0.2% glacial acetic acid solution; The mobile phase flow rate was 0.7~1.1 ml / min; Column temperature: 28~32℃; Detection wavelength: 270~290nm; The gradient elution procedure is as follows: At 0 minutes, mobile phase A is 5% and mobile phase B is 95%. At 10 minutes, mobile phase A was 5% and mobile phase B was 95%. At 45 minutes, mobile phase A was 19% and mobile phase B was 81%. At 60 minutes, mobile phase A was 45% and mobile phase B was 55%. At 65 minutes, mobile phase A was 50% and mobile phase B was 50%. At 79 minutes, mobile phase A was 50% and mobile phase B was 50%. At 85 minutes, mobile phase A was 70% and mobile phase B was 30%. At 105 minutes, mobile phase A was 72% and mobile phase B was 28%. At 110 minutes, mobile phase A was 5% and mobile phase B was 95%. At 120 minutes, mobile phase A was 5% and mobile phase B was 95%.

2. The method for constructing the fingerprint spectrum of Shengmai tea bags as described in claim 1, characterized in that, The methanol solution is a 50% aqueous methanol solution.

3. The method for constructing the fingerprint spectrum of Shengmai tea bags as described in claim 1, characterized in that, The flow rate of the mobile phase is 0.9 ml / min.

4. The method for constructing the fingerprint spectrum of Shengmai tea bags as described in claim 1, characterized in that, The column temperature is 30°C.

5. The method for constructing the fingerprint spectrum of Shengmai tea bags as described in claim 1, characterized in that, The detection wavelength is 275~285nm.

6. The method for constructing the fingerprint spectrum of Shengmai tea bags as described in claim 5, characterized in that, The detection wavelength is 280nm.

7. The method for constructing the fingerprint spectrum of Shengmai tea bags as described in claim 1, characterized in that, The detector is an ultraviolet detector.

8. The method for constructing the fingerprint spectrum of Shengmai tea bags as described in claim 1, characterized in that, The fingerprint spectrum of the Shengmai tea bags includes 15 marker peaks.

9. The method for constructing the fingerprint spectrum of Shengmai tea bags as described in claim 8, characterized in that, Of the 15 labeled peaks, peak 1 is 5-hydroxymethylfurfural, peak 5 is 4-coumaric acid, peak 8 is schisandrol A, peak 11 is schisandrol B, peak 14 is schisandrin A, and peak 15 is schisandrin B.

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