Method for detecting content of components in boenninghausenia sessilicarpa preparation
The characteristic map of the syrup preparation was constructed by high-performance liquid chromatography, which solved the problem that the content of rutin, isochrolactone and rutellide cannot be detected simultaneously in the prior art, and achieved an accurate and repeatable detection effect.
Patent Information
- Application Number
- CN202510742569.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-05
- Publication Date
- 2025-08-12
AI Technical Summary
The prior art lacks effective methods to simultaneously detect the ingredient content of rutin, isochrylactone and rutellide in the preparation of syrup.
High performance liquid chromatography was used to prepare test samples and reference solutions, and combined with chromatographic detection technology, a characteristic map of the preparation of rutin, isochrolactone and rutellide were constructed to detect the contents of rutin, isochrolactone and rutellide.
The accurate detection of the ingredient content in the preparation of shijiaocalyptus is achieved, the test results are stable and repeatable, and the testing process is simple.
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Figure CN120468342A_ABST
Abstract
Description
Technical Field
[0001] The present application relates to the technical field of detection methods, and in particular to a method for detecting the content of ingredients in a gypsophila preparation. Background Art
[0002] Peperomia belongs to the genus Peperomia of the Rutaceae family. It is mainly distributed in northeastern Yunnan and southwestern Sichuan, and is often found in mountainous areas with higher altitudes. It has the effects of dispelling wind and relieving exterior symptoms, clearing heat and detoxifying, promoting qi and activating blood circulation, and can be used to treat bronchitis, pneumonia, stomach pain and bloating, urinary tract infections, etc.
[0003] Currently, there are few reports on the detection of peperomia preparations, and there is a lack of methods for simultaneously detecting the content of rutin, isocarpin and rutin in peperomia preparations. Summary of the Invention
[0004] The purpose of this application is to provide a method for detecting the content of components in a peperomia preparation, which can effectively detect the component content and characteristic spectrum of rutin, isoprenolactone and rutin in the peperomia preparation.
[0005] The technical solution of this application is as follows:
[0006] The present invention provides a method for detecting the content of ingredients in a lycopodiella preparation, which comprises the following steps:
[0007] Preparation of test solution: Take the peperomia preparation, add solvent to perform ultrasonic extraction, and filter to obtain the test solution;
[0008] Preparation of reference solution: Take rutin, isoprenolide and rutinolide reference substances, mix them and add solvent to obtain reference solution;
[0009] Chromatographic detection: The reference solution and the test solution were respectively injected into the liquid chromatograph, and the determination was carried out by high performance liquid chromatography to obtain the characteristic spectrum of the phytoncide preparation and the contents of rutin, isocarpine and rutin in the test phytoncide preparation.
[0010] Furthermore, in some embodiments of the present application, the above-mentioned caltrop preparation is any one of caltrop slices, caltrop standard decoction, and caltrop formula granules.
[0011] Furthermore, in some embodiments of the present application, the solvent is methanol.
[0012] Furthermore, in some embodiments of the present application, in the above-mentioned test solution preparation step, 0.5 g of the lycopodiella preparation is taken, 10 ml of solvent is added, the weight is weighed, and then ultrasonic extraction is performed for 10 to 30 minutes. After cooling, the weight is weighed again. The previously weighed weight is used as the basis, and the lost weight is supplemented with the solvent, and then filtered to obtain the test solution.
[0013] Furthermore, in some embodiments of the present application, the rutin content in the above-mentioned reference solution is 0.3 mg / ml, the isoprenoids content is 0.15 mg / ml, and the rutin content is 0.15 mg / ml.
[0014] Furthermore, in some embodiments of the present application, in the above-mentioned chromatographic detection step, 5 μl of the reference solution and the test solution are respectively injected into a liquid chromatograph, and the determination is performed by high performance liquid chromatography.
[0015] Furthermore, in some embodiments of the present application, in the above-mentioned chromatography detection step, the liquid chromatography detection conditions are as follows:
[0016] The chromatographic column is filled with octadecylsilane bonded silica gel, the column temperature is 30-40°C, the mobile phase A is acetonitrile, the mobile phase B is 0.1% formic acid aqueous solution, the flow rate is 0.8-1 ml / min, the detection wavelength is 320 nm, and gradient elution is performed.
[0017] Furthermore, in some embodiments of the present application, in the above-mentioned chromatographic detection step, the gradient elution procedure is as follows:
[0018] From 0 to 30 min, the concentration of mobile phase A was increased from 10% (v / v) to 18.5% (v / v);
[0019] 30-40 min, mobile phase A was increased from 18.5% (v / v) to 21% (v / v);
[0020] At 40-45 min, the mobile phase A concentration was increased from 21% (v / v) to 29% (v / v);
[0021] From 45 to 67 min, the mobile phase A concentration was increased from 29% (v / v) to 36.5% (v / v);
[0022] From 67 to 72 min, the mobile phase A concentration increased from 36.5% (v / v) to 64% (v / v);
[0023] From 72 to 82 min, the mobile phase A concentration was increased from 64% (v / v) to 75% (v / v);
[0024] From 82 to 92 min, mobile phase A was maintained at 75% (v / v).
[0025] Compared with the prior art, the embodiments of the present application have at least the following advantages or beneficial effects:
[0026] In response to the above aspects, the embodiments of the present application provide a method for detecting the content of ingredients in a gypsophila preparation. The scheme adopts high-performance liquid chromatography, which can simultaneously detect the content of rutin, isocarpine and rutin contained in the gypsophila preparation and construct a characteristic spectrum; the detection method has a simple process, is easy to operate, and the detection results are stable and accurate with good repeatability. BRIEF DESCRIPTION OF THE DRAWINGS
[0027] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following is a brief introduction to the drawings required for use in the embodiments. It should be understood that the following drawings only show certain embodiments of the present application and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without creative work.
[0028] Figure 1 This is the characteristic spectrum of the standard decoction of Peperomia japonica in Example 1 of the present application;
[0029] Figure 2 This is the characteristic spectrum of the peperomia formula granules in Example 2 of this application;
[0030] Figure 3 This is the characteristic spectrum of the peperomia beverage in Example 3 of this application;
[0031] Figure 4 This is a graph showing the column temperature investigation results in Experimental Example 1 of this application;
[0032] Figure 5 This is a flow rate investigation result diagram in Test Example 1 of this application;
[0033] Figure 6 This is a diagram showing the results of solvent extraction in Experimental Example 1 of this application;
[0034] Figure 7 This is a graph showing the time investigation results extracted from Experimental Example 1 of this application;
[0035] Figure 8 This is the characteristic spectrum of the precision test in Test Example 2 of this application;
[0036] Figure 9 This is the characteristic spectrum of the repeatability inspection in Experimental Example 2 of this application;
[0037] Figure 10 This is the characteristic spectrum of durability investigation in Test Example 2 of this application. DETAILED DESCRIPTION
[0038] To make the purpose, technical solutions and advantages of the examples of the present application clearer, the technical solutions in the examples of the present application will be described clearly and completely below. Where specific conditions are not specified in the examples, conventional conditions or conditions recommended by the manufacturer are used. Where the manufacturer of the reagents or instruments is not specified, they are all conventional products that can be purchased commercially.
[0039] It should be noted that the term "comprises" or any other variation thereof is intended to cover non-exclusive inclusion, so that a process, method, article, or apparatus that includes a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article, or apparatus. In the absence of further limitations, the elements defined by the phrase "comprises..." do not exclude the presence of other identical elements in the process, method, article, or apparatus that includes the elements.
[0040] It should be noted that, unless there is any conflict, the embodiments and features in the embodiments of this application can be combined with each other.
[0041] The features and performance of the present application are further described in detail below with reference to the embodiments.
[0042] Example 1
[0043] This embodiment provides a method for detecting the content of components in a lycopodium preparation, and the method steps are as follows:
[0044] Preparation of test solution: Take 0.5 g of standard decoction of Peperomia japonica, add 10 ml of methanol, weigh the weight, extract by ultrasonic for 10 min, cool and weigh again, use the previously weighed weight as the basis, make up the lost weight with methanol, shake well and filter, take the filtrate to obtain the test solution.
[0045] Preparation of reference solution: Take rutin, isoprenaline and rutin lactone reference substances, mix them and add solvent to obtain a reference solution. The reference solution has a rutin content of 0.3 mg / ml, an isoprenaline content of 0.15 mg / ml, and a rutin content of 0.15 mg / ml.
[0046] Chromatographic detection: Accurately aspirate 5 μl of the reference solution and the test solution, inject them into a liquid chromatograph, and determine them by high performance liquid chromatography to obtain the characteristic spectrum of the phytoncide preparation and the contents of rutin, isocarpine, and rutin in the test phytoncide preparation;
[0047] Liquid chromatography conditions are as follows:
[0048] Chromatographic column: Octadecylsilane bonded silica gel was used as filler (Chromatographic column Poroshell 120SB-C18, 4.6 mm × 250 mm, 4 μm);
[0049] Column temperature: 30°C;
[0050] Mobile phase A: acetonitrile;
[0051] Mobile phase B: 0.1% formic acid in water;
[0052] Flow rate: 1 ml / min;
[0053] Detection wavelength: 320nm;
[0054] The gradient elution program is as follows:
[0055] From 0 to 30 min, the concentration of mobile phase A was increased from 10% (v / v) to 18.5% (v / v);
[0056] 30-40 min, mobile phase A was increased from 18.5% (v / v) to 21% (v / v);
[0057] At 40-45 min, the mobile phase A concentration was increased from 21% (v / v) to 29% (v / v);
[0058] From 45 to 67 min, the mobile phase A concentration was increased from 29% (v / v) to 36.5% (v / v);
[0059] From 67 to 72 min, the mobile phase A concentration increased from 36.5% (v / v) to 64% (v / v);
[0060] From 72 to 82 min, the mobile phase A concentration was increased from 64% (v / v) to 75% (v / v);
[0061] From 82 to 92 min, mobile phase A was maintained at 75% (v / v).
[0062] According to the above method, 15 batches of standard decoction of Peperomia were tested, and the results are as follows:
[0063] The contents of rutin, isocarpin and rutin in 15 batches of standard decoctions of caltrop were tested, and the results are shown in Table 1:
[0064] Table 1
[0065]
[0066]
[0067] As shown in Table 1, the content of the index component rutin in the 15 batches of standard decoctions ranged from 1.35 to 9.69 mg / g, with an average of 4.3 mg / g, a SD of 249.5%, and an RSD of 58.04%. The 70% to 130% range of the average rutin content was 3.01 to 5.59 mg / g; the 70% to 130% range of the average isoanisole was 0.21 to 0.72 mg / g, with an average of 0.42 mg / g, a SD of 17.43%, and an RSD of 41.21%. The 70% to 130% range of the average isoanisole was 0.3 to 0.55 mg / g; and the rutin content ranged from 0.14 to 0.28 mg / g, with an average of 0.19 mg / g, a SD of 4.11%, and an RSD of 21.55%. The range of 70% to 130% of the average value of the rutin content is 0.13 to 0.25 mg / g.
[0068] The relative retention time ratios of 15 batches of standard decoctions of Peperomia japonica are shown in Table 2:
[0069] Table 2
[0070]
[0071] The relative peak areas of 15 batches of standard decoctions of Peperomia japonica are shown in Table 3:
[0072] Table 3
[0073]
[0074]
[0075] Combining Tables 2 and 3, five peaks with good reproducibility were selected as characteristic peaks based on the principles of stable relative retention times, consistent detection across all batches of samples, and relatively high peak heights. The results showed that when Peak 1 was used as the S peak, the relative peak area RSDs for the characteristic peaks in all 15 batches of Peperomia standard decoctions ranged from 61.29% to 101.79%, and the relative retention time RSDs for the five characteristic peaks in all 15 batches of Peperomia standard decoctions were all less than 0.77%.
[0076] The summary standard of RSD (%) of each item result of the methodology (relative retention time) is shown in Table 4:
[0077] Table 4
[0078]
[0079] The summary standard of RSD (%) of each item result of the methodology (relative peak area) is shown in Table 5:
[0080] Table 5
[0081]
[0082] Final regulations: The test sample's characteristic spectrum should show five characteristic peaks. The first peak, S, should be identical to the rutin reference peak. The fourth peak should be identical to the isothiocyanate peak. The fifth peak should be identical to the rutin peak. Calculate the relative retention time of each characteristic peak relative to the S peak. The relative retention time should be within ±8% of the specified value. The specified values are: 1.7960 (peak 2), 2.1085 (peak 3), 2.2188 (peak 4), and 2.9058 (peak 5).
[0083] like Figure 1 As shown in the figure, 15 batches of standard decoctions of Herba Piperis were synthesized using the Chinese medicine chromatographic fingerprint similarity evaluation system (2012 version), and a comparison map of the characteristic spectra of the standard decoctions of Herba Piperis was established.
[0084] Example 2
[0085] This example is based on Example 1, and differs from Example 1 in that, in the steps of preparing the test solution, the standard decoction of caltrops is replaced with caltrops formula granules, and the remaining methods and conditions are the same as those of Example 1.
[0086] Three batches of peperomia formula granules were tested, and the results are as follows:
[0087] The test results of rutin, isocarpine and rutin in three batches of physalis formula granules are shown in Table 6:
[0088] Table 6
[0089]
[0090] The relative retention time ratios of the three batches of physalis formula granules are shown in Table 7:
[0091] Table 7
[0092]
[0093] The relative peak areas of three batches of physalis formula granules are shown in Table 8:
[0094] Table 8
[0095]
[0096]
[0097] Based on the principles of stable relative retention times, consistent detection across all batches of samples, and relatively high peaks, five peaks with good reproducibility were selected as characteristic peaks. The results showed that when Peak 1 was used as the S peak, the RSDs of the relative retention times of the five characteristic peaks in the three batches of Peperomia formula granules were all less than 2.53%.
[0098] like Figure 2As shown in the figure, three batches of Herba Cibotii formula granules were synthesized using the Chinese medicine chromatographic fingerprint similarity evaluation system (2012 version), and a comparison map of the characteristic spectrum of Herba Cibotii formula granules was established.
[0099] Example 3
[0100] This embodiment is based on Example 1, and differs from Example 1 in that, in the above-mentioned test solution preparation step, the standard decoction of Peperomia japonica is replaced with Peperomia japonica slices, and the remaining methods and conditions are the same as those in Example 1.
[0101] Fifteen batches of Peperomia japonica slices were tested, and the results are as follows:
[0102] The test results of rutin, isocarpine and rutin in 15 batches of caltrop slices are shown in Table 9:
[0103] Table 9
[0104]
[0105] The content of the index component rutin in 15 batches of gypsophila slices ranged from 0.77 to 7.82 mg / g, with an average of 2.48 mg / g, a SD of 218.38%, and an RSD of 87.93%. The 70% to 130% range of the average rutin content was 1.74 to 3.23 mg / g. The content of isoanisole lactone ranged from 0.10 to 0.67 mg / g, with an average of 0.44 mg / g, a SD of 16.1%, and an RSD of 36.52%. The 70% to 130% range of the average isoanisole lactone content was 0.13 to 0.57 mg / g. The content of rutin ranged from 0.77 to 2.58 mg / g, with an average of 1.78 mg / g, a SD of 45.38%, and an RSD of 25.42%. The range of 70% to 130% of the average value of the rutin content is 1.25 to 2.32 mg / g.
[0106] The relative retention time ratios of 15 batches of Peperomia slices are shown in Table 10
[0107] Table 10
[0108]
[0109] The relative peak areas of 15 batches of caltrop beverages are shown in Table 11:
[0110] Table 11
[0111]
[0112]
[0113] Based on the principles of stable relative retention times, consistent detection across all batches, and relatively high peak heights, five peaks with good reproducibility were selected as characteristic peaks. The results showed that when Peak 1 was used as the S peak, the relative peak area RSDs of the characteristic peaks for all 15 batches of Peperomia japonica slices were too large and, therefore, were not included in the quality standard. The relative retention time RSDs of the characteristic peaks were all less than 0.83%.
[0114] like Figure 3 As shown in the figure, 15 batches of Herba Cyperi decoction pieces were synthesized using the Chinese medicine chromatographic fingerprint similarity evaluation system (2012 version), and a comparison map of the characteristic spectra of Herba Cyperi decoction pieces was established.
[0115] Test Example 1
[0116] Based on the detection method in Example 1, the method conditions were investigated as follows:
[0117] 1. Column temperature inspection
[0118] Based on the conditions of the method in Example 1, investigations were conducted at column temperatures of 30°C, 35°C, and 40°C.
[0119] like Figure 4 As shown in the figure, S1, S2, and S3 refer to 30°C, 35°C, and 40°C, respectively. The results show that when the column temperature is 30°C, the chromatogram peaks are relatively symmetrical and the separation is good, so the column temperature is set to 30°C.
[0120] 2. Flow rate investigation
[0121] Based on the conditions of the method in Example 1, investigations were conducted at flow rates of 0.8 mL / min, 1.0 mL / min, and 1.2 mL / min.
[0122] like Figure 5 As shown in the figure, S1, S2, and S3 represent 0.8 mL / min, 1.0 mL / min, and 1.2 mL / min, respectively. The results show that at a flow rate of 1.0 mL / min, the chromatogram has good peak shape and moderate resolution. Therefore, the flow rate was determined to be 1.0 mL / min.
[0123] 3. Extraction solvent investigation
[0124] Take 0.5 g of sample (batch number 2301222), accurately weigh it, and place it in a stoppered conical flask. The extraction solvents for the test sample were methanol, 70% methanol, and 50% methanol, 10 mL each. Seal the flask, weigh it, and ultrasonically treat it (power 600 W, frequency 40 kHz) for 10 minutes. Let it cool, weigh it again, make up the lost weight with the extraction solvent, shake it well, filter it, and take the filtrate.
[0125] like Figure 6As shown in the figure, S1, S2, and S3 refer to methanol, 70% methanol, and 50% methanol, respectively. The results show that when methanol is the extraction solvent, the chromatographic peaks are more informative and the baseline is more stable. Therefore, methanol was determined to be the extraction solvent for the test sample.
[0126] 4. Extraction time investigation
[0127] Take 0.5 g of sample (batch number 2301222), accurately weigh it, place it in a stoppered conical flask, accurately add 25 mL of methanol, stopper it, weigh it, and treat it ultrasonically (power 600 W, frequency 40 kHz). The extraction time of the test sample was 10 minutes, 20 minutes, and 30 minutes respectively. Let it cool, weigh it again, make up the lost weight with methanol, shake it well, filter it, and take the filtrate.
[0128] like Figure 7 As shown in the figure, S1, S2, and S3 represent 10 minutes, 20 minutes, and 30 minutes, respectively. The results show that complete extraction can be achieved when the extraction time is 10 minutes to 30 minutes. Considering all factors, the extraction time is set to 10 minutes.
[0129] Test Example 2
[0130] A methodological investigation was conducted on the method of Example 1.
[0131] 1. Chromatographic peak identification
[0132] According to the method of Example 1, a test solution of a standard decoction of gypsophila and a reference solution were prepared;
[0133] The characteristic chromatographic peaks of the standard decoction of Peperomia japonica were identified.
[0134] like Figure 1 As shown, the results show that peak 1 is rutin, peak 4 is isocarpine, and peak 5 is rutin. In the following methodological investigation, the five peaks in the sample were investigated;
[0135] In the following methodological investigation, five peaks in the sample were investigated.
[0136] 2. Precision test
[0137] Take the test solution of Peperomia japonica standard decoction (batch number 2301222) and inject it continuously 6 times according to the proposed experimental method, 5 μL each time, and calculate the retention time and peak area of each characteristic peak.
[0138] The results are shown in Table 12 (retention time) and Table 13 (peak area); its characteristic spectrum is shown in Figure 8 shown.
[0139] Table 12
[0140]
[0141] Table 13
[0142]
[0143] The results showed that the RSD values of the retention time and peak area of each characteristic peak met the requirements and had good precision.
[0144] 3. Repeatability test
[0145] Six portions of freeze-dried powder of standard decoction of Peperomia japonica (batch number 2301222) were accurately weighed and prepared and determined according to the proposed experimental method. The relative retention time and relative peak area of each characteristic peak were calculated.
[0146] The results are shown in Table 15 (relative retention time) and Table 16 (relative peak area); its characteristic spectrum is shown in Figure 9 shown.
[0147] Table 15
[0148]
[0149] Table 16
[0150]
[0151] The results showed that the RSD values of the retention time and peak area of each characteristic peak met the requirements and had good repeatability.
[0152] 4. Durability inspection
[0153] Based on the conditions of the method in Example 1, the same test solution was measured at 0 h, 2 h, 4 h, 8 h, 18 h, 12 h, and 24 h. The retention time and peak area of each characteristic peak were calculated.
[0154] The results are shown in Table 17 (retention time) and Table 18 (peak area); its characteristic spectrum is shown in Figure 10 shown.
[0155] Table 17
[0156]
[0157] Table 18
[0158]
[0159] The results showed that the RSD of the corresponding characteristic peak retention time was between 0.07% and 0.51%, and the characteristic peak area was between 0.00% and 5.94%. The sample solution was relatively stable within 24 hours.
[0160] In summary, the embodiments of the present application provide a method for detecting the content of ingredients in a peperomia preparation. The scheme adopts high-performance liquid chromatography, which can simultaneously detect the content of rutin, isoprenaline and rutin contained in the peperomia preparation and construct a characteristic spectrum; the detection method is simple in process, easy to operate, and the detection results are stable, accurate and reproducible.
[0161] The embodiments described above are part of the embodiments of the present application, rather than all of the embodiments. The detailed description of the embodiments of the present application is not intended to limit the scope of the present application for protection, but merely represents selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.
Claims
1. A method for detecting the content of ingredients in a gypsophila preparation, characterized in that: It includes the following steps: Preparation of test solution: Prepare the test solution using the caltrop preparation; Preparation of reference solution: Take rutin, isoprenolide and rutinolide reference substances, mix them and add solvent to obtain reference solution; Chromatographic detection: The reference solution and the test solution were respectively injected into the liquid chromatograph, and the determination was carried out by high performance liquid chromatography to obtain the characteristic spectrum of the phytoncide preparation and the contents of rutin, isocarpine and rutin in the test phytoncide preparation.
2. The method for detecting the content of components in a lycopodii preparation according to claim 1, wherein: The caltrop preparation is any one of caltrop slices, caltrop standard decoction, and caltrop formula granules.
3. The method for detecting the content of components in a lycopodii preparation according to claim 1, wherein: The test solution is prepared by taking a peperomia preparation, adding a solvent to perform ultrasonic extraction, and filtering to obtain the test solution.
4. The method for detecting the content of components in a lycopodii preparation according to claim 3, characterized in that: The solvent is methanol.
5. The method for detecting the content of components in a lycopodii preparation according to claim 1, wherein: In the test solution preparation step, the gypsophila preparation is first taken and a solvent is added thereto, and then the weight is weighed. Then, ultrasonic extraction is performed, and the weight is weighed again after cooling. The previously weighed weight is used as a reference, and the lost weight is supplemented with the solvent. Then, the test solution is filtered to obtain the test solution.
6. The method for detecting the content of components in a lycopodii preparation according to claim 5, characterized in that: In the test solution preparation step, 0.5 g of the gypsophila preparation is taken, 10 ml of solvent is added, and the weight is weighed. Then, ultrasonic extraction is performed for 10 to 30 minutes. After cooling, the weight is weighed again. The previously weighed weight is used as the basis, and the lost weight is supplemented with the solvent. Then, the test solution is filtered to obtain the test solution.
7. The method for detecting the content of components in a lycopodii preparation according to claim 1, characterized in that: The reference solution contained 0.3 mg / ml of rutin, 0.15 mg / ml of isoprenolide, and 0.15 mg / ml of rutin.
8. The method for detecting the content of components in a lycopodii preparation according to claim 1, characterized in that: In the chromatographic detection step, 5 μl of the reference solution and the test solution were respectively injected into a liquid chromatograph and measured by high performance liquid chromatography.
9. The method for detecting the content of components in a lycopodii preparation according to claim 1, characterized in that: In the chromatographic detection step, the liquid chromatography detection conditions are as follows: The chromatographic column is filled with octadecylsilane bonded silica gel, the column temperature is 30-40°C, the mobile phase A is acetonitrile, the mobile phase B is 0.1% formic acid aqueous solution, the flow rate is 0.8-1 ml / min, the detection wavelength is 320 nm, and gradient elution is performed.
10. The method for detecting the content of components in a lycopodii preparation according to claim 9, characterized in that: In the chromatographic detection step, the gradient elution procedure is as follows: From 0 to 30 min, the concentration of mobile phase A was increased from 10% (v / v) to 18.5% (v / v); 30-40 min, mobile phase A was increased from 18.5% (v / v) to 21% (v / v); At 40-45 min, the mobile phase A concentration was increased from 21% (v / v) to 29% (v / v); From 45 to 67 min, the mobile phase A concentration was increased from 29% (v / v) to 36.5% (v / v); From 67 to 72 min, the mobile phase A concentration increased from 36.5% (v / v) to 64% (v / v); From 72 to 82 min, the mobile phase A concentration was increased from 64% (v / v) to 75% (v / v); From 82 to 92 min, mobile phase A was maintained at 75% (v / v).