Characteristic spectrum detection method of Fuyinkang lotion

Abstract

The invention provides a specific chromatogram detection method of Fuyinkang lotion, and belongs to the technical field of pharmaceutical preparation detection. According to the specific chromatogram detection method, high performance liquid chromatography is adopted for detection, and a chromatographic column of the specific chromatogram detection method is a chromatographic column with octadecylsilane chemically bonded silica as a filler; a mobile phase is acetonitrile and a 0.1% acidic aqueous solution; the column temperature is 29 DEG C to 31 DEG C, and the flow velocity is 0.98 mL / min to 1.02 mL / min; the detection wavelength is 220nm. According to the specific chromatogram detection method provided by the invention, the specific chromatogram with more stable base lines and more characteristic peaks can be obtained, the separation effect of each characteristic peak is better, and more known component types are contained. The characteristic spectrum can comprehensively reflect characteristic peak information of the Fuyinkang lotion, is suitable for overall quality control of the Fuyinkang lotion, and can comprehensively and effectively evaluate the quality of the Fuyinkang lotion.

Description

Technical Field

[0001] The invention belongs to the technical field of drug preparation detection, and particularly relates to a characteristic spectrum detection method for Fuyinkang lotion. Background Art

[0002] Fuyinkang wash is composed of 6 Chinese medicinal herbs, namely, Qinpi, Houttuynia cordata, Sophora flavescens, Agrimoniae herba, Isatis indigotica and Viola yedoensis. It is prepared by decocting with water, concentrating, precipitating with alcohol, concentrating, preparing liquid, filling and sterilizing. It has the effects of clearing away heat and dampness, relieving itching and stopping leukorrhea. It is mainly used for bacterial vaginosis, Trichomonas vaginitis, Candida vaginitis, and those who are diagnosed with damp-heat accumulation according to TCM syndrome. It can improve the symptoms of vulvar itching and pain, excessive leukorrhea, frequent urination, urgency and pain in urination.

[0003] The medicinal material Qinpi mainly contains coumarin compounds such as qinpi acetone, qinpi ethyl, qinpi glycoside, and qinpi acetone. The Chinese Pharmacopoeia stipulates that the base sources of Qinpi medicinal materials include 4 species of the genus Fraxinus in the family Oleaceae. Because Qinpi is a multi-source variety, and the geographical environment of the original plant distribution is different, the quality of commercial medicinal materials from different origins and sources varies greatly. At the same time, the types of Qinpi and mixed medicinal materials are also relatively complex, which needs to be clarified urgently. At present, the internal quality control of Qinpi mostly adopts the content determination method of qinpi acetone and qinpi ethyl. The journal literature ("Research on the Improvement of the Production Process of Fuyinkang Lotion", Tang Yingwu et al., "Journal of Kaili College", Issue 6, 2021) records that the current quality standard of Fuyinkang Lotion mainly evaluates and controls its quality by controlling the content of a single ingredient qinpi ethyl, which has certain limitations.

[0004] Therefore, in order to better reflect the intrinsic quality of Fuyinkang Lotion, it is very necessary to establish a characteristic spectrum detection method and quality control method for Fuyinkang Lotion. Summary of the invention

[0005] Based on the deficiencies in the prior art, the present invention discloses a gynecological external Chinese medicine composition, namely, Fuyinkang lotion, with Chinese patent number ZL202311736809.6, and provides a characteristic spectrum detection method. The characteristic spectrum provided by the present invention can comprehensively and effectively evaluate the quality of Fuyinkang lotion.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] A characteristic spectrum detection method for Fuyinkang lotion, wherein the characteristic spectrum detection method adopts high performance liquid chromatography to detect and obtain the characteristic spectrum of Fuyinkang lotion;

[0008] The chromatographic column used in the characteristic spectrum detection method is a chromatographic column with octadecylsilane bonded silica gel as a filler;

[0009] The characteristic spectrum detection method adopts acetonitrile and 0.1% acidic aqueous solution as mobile phase; the acidic aqueous solution is phosphoric acid aqueous solution or formic acid aqueous solution.

[0010] The elution of the characteristic spectrum detection method is gradient elution;

[0011] The gradient elution is as follows: acetonitrile is used as mobile phase A, 0.1% phosphoric acid aqueous solution is used as mobile phase B, and elution is performed according to the following gradient elution program:

[0012]

[0013] The detection wavelength of the characteristic spectrum detection method is 220nm.

[0014] In the characteristic spectrum detection method, the column temperature of the chromatographic column is 29° C.-31° C., and the flow rate is 0.98 mL / min-1.02 mL / min.

[0015] Preferably, the chromatographic column is Kromasil 100-5-C18.

[0016] The preparation process of the test solution in the characteristic spectrum detection method is: take Fuyinkang lotion, add solvent, shake well, and filter to obtain the filtrate which is the test solution.

[0017] The solvent is methanol or water, preferably water.

[0018] The standard characteristic spectrum has 9 characteristic peaks, and the peak areas account for more than 90% of the total peak area; the chromatographic peak No. 3 is the characteristic peak of aesculin, and the relative retention time and relative peak area of ​​the characteristic peak of aesculin are 1. The relative retention time and relative peak area of ​​other peaks are calculated to obtain the standard characteristic spectrum of Fuyinkang lotion;

[0019] The characteristic spectrum of the Fuyinkang lotion includes characteristic peaks corresponding to matrine, chlorogenic acid, aesculetin, fraxinoside, and akebia phenylethanoid glycoside B, wherein matrine is peak 1, aesculetin is peak 3, chlorogenic acid is peak 4, aesculetin is peak 5, fraxinoside is peak 6, and akebia phenylethanoid glycoside B is peak 8; peak 3 is peak S, and the retention time of other characteristic peaks relative to peak 3 is ±10% of the specified value, and the specified value of the characteristic peak is:

[0020] Peak 1: 0.45, Peak 2: 0.73, Peak 4: 1.30, Peak 5: 1.45, Peak 6: 1.49, Peak 7: 1.84, Peak 8: 1.97, Peak 9: 2.74.

[0021] Taking Peak 3 as the S peak, the relative peak area of ​​Peak 6 shall not be less than 0.34.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] The present invention provides a characteristic spectrum detection method for Fuyinkang lotion, by selecting aesculin as an internal reference, whose relative retention time is 1.00, and then determining that the retention time of other characteristic peaks in the characteristic spectrum relative to aesculin is ±10% of a specified value, and the specified values ​​of the characteristic peaks are: peak 1: 0.45, peak 2: 0.73, peak 4: 1.30, peak 5: 1.45, peak 6: 1.49, peak 7: 1.84, peak 8: 1.97, peak 9: 2.74; so that the obtained characteristic spectrum can comprehensively and effectively evaluate the quality of Fuyinkang lotion.

[0024] The characteristic spectrum detection method of the Fuyinkang lotion of the present invention can obtain a characteristic spectrum with a more stable baseline and more characteristic peaks, and the separation effect of each characteristic peak is better, and more types of known components are included. Therefore, the characteristic spectrum can fully reflect the characteristic peak information of the Fuyinkang lotion, is suitable for the overall quality control of the Fuyinkang lotion, and can comprehensively and effectively evaluate the quality of the Fuyinkang lotion. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0026] Figure 1 It is the comparative characteristic spectrum of the Fuyinkang lotion in the present invention.

[0027] Figure 2 It is the positioning characteristic spectrum of matrine, aesculin, chlorogenic acid, aesculetin, aesculin, akebia phenylethanoid glycoside B and Fuyinkang lotion in the present invention.

[0028] Figure 3 It is the characteristic spectrum of the Fuyinkang lotion in the present invention detected at 220nm.

[0029] Figure 4 It is the characteristic spectrum of the Fuyinkang lotion in the present invention detected at 254nm.

[0030] Figure 5 It is the characteristic spectrum of the Fuyinkang lotion in the present invention detected at 280nm.

[0031] Figure 6 It is the characteristic spectrum of the Fuyinkang lotion in the present invention detected at 320nm.

[0032] Figure 7It is the characteristic spectrum of the Fuyinkang lotion in the present invention when eluted with 0.1% phosphoric acid aqueous solution as mobile phase B.

[0033] Figure 8 It is the characteristic spectrum of the Fuyinkang lotion in the present invention when eluted with 0.1% formic acid aqueous solution as mobile phase B.

[0034] Fig. 9 This is the characteristic spectrum of the Fuyinkang lotion in the present invention detected by the chromatographic column 1.

[0035] Fig.10 This is the characteristic spectrum of the Fuyinkang lotion in the present invention detected by chromatographic column 2.

[0036] Fig.11 This is the characteristic spectrum of the Fuyinkang lotion in the present invention detected by chromatographic column 3.

[0037] Fig.12 This is the characteristic spectrum obtained by using the gradient elution program of Table 1 for the Fuyinkang lotion of the present invention.

[0038] Fig.13 This is the characteristic spectrum obtained by using the gradient elution program of Table 8 for the Fuyinkang lotion of the present invention.

[0039] Fig.14 This is the characteristic spectrum obtained when the Fuyinkang lotion of the present invention uses water as the test sample solvent.

[0040] Fig.15 This is the characteristic spectrum obtained when the Fuyinkang lotion of the present invention uses methanol as the test sample solvent.

[0041] Fig.16 It is the test spectrum of Fuyinkang lotion under the prior art. DETAILED DESCRIPTION

[0042] The present invention discloses a characteristic spectrum detection method for Fuyinkang lotion, and those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It should be particularly pointed out that all similar substitutions and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention.

[0043] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the literature in the art can be used. If no manufacturer is specified for the reagents or instruments used, they are all conventional reagent products that can be obtained commercially. The percentages not specified in the present invention are all volume percentages.

[0044] instrument:

[0045] Chromatograph 1: Agilent 1260Ⅱ chromatography system, including G7111B quaternary pump, G7129A autosampler, G7114A diode array detector, and chromatography workstation.

[0046] Chromatograph 2: UltiMate 3000 chromatography system, including LPG-3400A quaternary pump, WPS-3000TSL autosampler, PDA diode array detector, chromatography workstation.

[0047] M2020 1 / 100,000 analytical balance (Mettler-Toledo), KQ-500DE ultrasonic instrument (Kunshan Ultrasonic Instrument Co., Ltd.)

[0048] Drug testing:

[0049] Matrine (batch number 110805-202010, for content determination, calculated as 98.7%) was purchased from China Food and Drug Administration

[0050] Aesculin (batch number 110740-201806, for content determination, calculated as 92.4%) was purchased from the China Food and Drug Administration

[0051] Chlorogenic acid (batch number 110753-202119, for content determination, calculated as 96.3%) was purchased from China Food and Drug Administration

[0052] Esculetin (batch number 110741-202310, for content determination, calculated as 98.3%) was purchased from China Food and Drug Administration

[0053] Fraxinoside (batch number ST00130120, for content determination, calculated as 98.4%) was purchased from Shanghai Shidande Standard Technology Service Co., Ltd.

[0054] Akebia phenylethanol glycoside B (batch number RS07141120, for content determination, calculated as 91.1%) was purchased from Shanghai Standard Standard Technology Service Co., Ltd.

[0055] Fuyinkang lotion (batch numbers: 221011, 221105, 230105, 230106, 231201, 231209, 240401, 240403, 240404, 240701) are all produced by Suzhou Yifan Pharmaceutical Co., Ltd.

[0056] Reagents: Acetonitrile was of chromatographic grade, water was ultrapure water, and other reagents were of analytical grade.

[0057] Example 1 A characteristic spectrum detection method for Fuyinkang lotion

[0058] High performance liquid chromatography (HPLC) was used for detection. The chromatographic conditions of the high performance liquid chromatography were as follows:

[0059] Chromatographic column: Octadecylsilane bonded silica gel as filler (Kromasil 100-5-C18, specification 4.6×250 mm, particle size 5 μm); acetonitrile as mobile phase A, 0.1% phosphoric acid aqueous solution as mobile phase B, elution according to the gradient elution program specified in Table 1; detection wavelength 220 nm; column temperature 30°C; flow rate 1.0 mL / min.

[0060] Table 1

[0061] Time (min) Flow rate (ml / min) Mobile phase A (%) Mobile phase B (%) 0-40 1.0 5→20 95→80 40-60 1.0 20→40 80→60

[0062] Preparation of reference solution: Take appropriate amount of matrine, fraxinoyl, fraxinoyl, chlorogenic acid, fraxinoside, and akebia phenylethanol glycoside B reference substances, weigh them accurately, and add methanol to make a mixed reference solution containing 200 μg of matrine, 80 μg of fraxinoyl, 40 μg of fraxinoyl, 40 μg of chlorogenic acid, 60 μg of fraxinoside, and 20 μg of akebia phenylethanol glycoside B per 1 mL.

[0063] Preparation of the test solution: Accurately measure 5 mL of Fuyinkang Lotion, place it in a 25 mL volumetric flask, add water to make up to volume, shake well, filter and take the filtrate.

[0064] Determination method: Accurately pipette 5 μL of reference solution and 10 μL of test solution respectively, inject into liquid chromatograph, and determine.

[0065] In this embodiment, characteristic spectra generated by 10 batches of Fuyinkang lotion were used, and the fingerprint similarity evaluation software "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System 2012 Edition" compiled by the State Pharmacopoeia Committee was used. The "multi-point correction, MARK peak matching" mode was used for fitting and generating a control characteristic spectrum to obtain the control characteristic spectrum of Fuyinkang lotion; after chromatographic matching, 9 common characteristic peaks with more appropriate response values, better separation and better purity were obtained, and the chromatographic peak numbers were rearranged as peak 1 to peak 9 according to the order of the chromatographic peaks, such as Figure 1 shown.

[0066] The characteristic peaks of the characteristic spectrum of Fuyinkang lotion were analyzed by high performance liquid chromatography-high resolution mass spectrometry. According to the multi-level mass spectrometry information of the sample, combined with the natural product high resolution mass spectrometry database and related literature, and HPLC was used to locate the reference substance. The specific relevant results are as follows Figure 2 And as shown in Table 2.

[0067] Table 2

[0068]

[0069] Note: “a” was verified by reference substance; “-” was not identified.

[0070] According to the above results, peak 1 was determined to be matrine, peak 3 was aesculin, peak 4 was chlorogenic acid, peak 5 was aesculin, peak 6 was aesculin, and peak 9 was akebia phenylethanoid glycoside B. Aesculin had good response value and separation, so aesculin was used as a reference peak to better evaluate the relative retention time and relative peak area of ​​each characteristic peak.

[0071] Based on the characteristic spectra generated by the above 10 batches of Fuyinkang lotion, the relative retention time and relative peak area of ​​each characteristic peak were calculated with aesculin as the S peak, as shown in Tables 3 and 4 below.

[0072] Table 3

[0073]

[0074]

[0075] As shown in Table 3, there should be 9 characteristic peaks in the chromatogram of the test sample. The relative retention time of each characteristic peak and the S peak should be calculated, and the relative retention time should be within ±10% of the specified value. The specified values ​​are: 0.45 (peak 1), 0.73 (peak 2), 1.30 (peak 4), 1.45 (peak 5), 1.49 (peak 6), 1.84 (peak 7), 1.97 (peak 8), 2.74 (peak 9).

[0076] Table 4

[0077]

[0078]

[0079] As shown in Table 4, the relative peak areas of peaks 4, 5, 6 and 9 calculated with fraxin as the reference peak have small deviations between batches, indicating that the differences between peaks 4, 5, 6 and 9 are small. Peak 6 has better stability between batches and a moderate peak area. The characteristic peaks of the characteristic spectrum of Fuyinkang lotion and the chromatographic peaks of fraxinoside reference substance were analyzed by high performance liquid chromatography-high resolution mass spectrometry. According to the multi-level mass spectrometry information of the sample and the reference substance, combined with the natural product resolution mass spectrometry database and related literature, peak 6 was determined to be fraxinoside; therefore, the relative peak area of ​​peak 6 was controlled to improve the quality control level of Fuyinkang lotion. Taking the chromatographic peak of qinpijuncin as the reference peak (S peak), the relative peak areas of peak 6 and S peak were calculated. The measured range of the relative peak areas of peak 6 in the characteristic spectra of 10 batches of Fuyinkang lotion samples was 0.43-0.55, with a mean of 0.49 and a SD of 0.03. Therefore, the relative peak area value of peak 6 is stipulated to be not less than 0.34 based on the mean of -30%. The quality control of Fuyinkang lotion can be achieved through the relative retention time and the relative peak area value of peak 6.

[0080] Example 2

[0081] The difference from Example 1 is that the chromatographic conditions are different, as follows:

[0082] 1. Detection wavelength

[0083] The present invention uses DAD detector to examine the HPLC graphs of samples at 220nm, 254nm, 280nm and 320nm respectively. The detection results are as follows: Figure 3-6 As shown; the investigation results show that at a wavelength of 220-280nm, with the increase of wavelength, the response values ​​of some characteristic peaks gradually increase, but at the same time, the peak information amount decreases with the increase of wavelength. At a wavelength of 220nm, the baseline of the characteristic peak is stable, the peak information amount is large, and the responses of each peak are relatively balanced. Taking comprehensive considerations, 220nm is selected as the detection wavelength of the characteristic spectrum of Fuyinkang Lotion.

[0084] 2. Flow rate

[0085] Take Fuyinkang lotion (batch number: 240403) and prepare the test solution according to the preparation method of the test solution in Example 1. Different flow rates (0.98 ml / min, 1.0 ml / min, 1.02 ml / min) were used and the other conditions were the same as in Example 1. The results are shown in Tables 5 to 7 below.

[0086] Table 5

[0087]

[0088] Table 6

[0089]

[0090]

[0091] Table 7

[0092]

[0093] The peaks in Tables 5 to 7 above are characteristic peaks marked corresponding to the characteristic peaks in the control spectrum in Example 1. The results of the investigation at different flow rates show that different flow rates (0.98 ml / min, 1.00 ml / min, 1.02 ml / min) have a certain influence on the theoretical plate number and separation degree of each peak, but the results of the flow rate detection within the above range all meet the system suitability requirements, preferably 1.0 ml / min.

[0094] 3. Mobile phase

[0095] The present invention uses a DAD detector to examine the HPLC graphs of samples eluted with 0.1% phosphoric acid aqueous solution and 0.1% formic acid aqueous solution as mobile phase B. The detection results are as follows: Figure 7 and Figure 8 As shown; a comparative study was conducted on mobile phase phosphoric acid and formic acid. The results showed that under the elution condition of 0.1% phosphoric acid aqueous solution as mobile phase B, the baseline of the characteristic peak was more stable and the separation was better. Taking comprehensive considerations, acetonitrile-0.1% phosphoric acid aqueous solution was selected as the mobile phase for the characteristic spectrum of Fuyinkang Lotion.

[0096] 4. Types of chromatographic columns

[0097] Take Fuyinkang lotion (batch number: 240403), prepare a test solution according to the preparation method of the test solution in Example 1, use the gradient elution in Example 1 and use different types of chromatographic columns to measure under the same other conditions as in Example 1. The results are as follows: Figure 9-11 shown.

[0098] Column 1: Hypersil GOLD-C18 (5 μm, 4.6 mm*250 mm)

[0099] Column 2: Inertil ODS-3C18 (5μm, 4.6mm*250mm)

[0100] Column 3: Kromasil 100-5-C18 (5 μm, 4.6 mm*250 mm)

[0101] Through the above Figure 9-11It can be seen that when using the Kromasil 100-5-C18 chromatographic column, the chromatographic peaks have richer information and better separation, so the chromatographic column is preferably the Kromasil 100-5-C18 chromatographic column.

[0102] 5. Elution gradient

[0103] Take Fuyinkang lotion (batch number: 240403) and prepare a test solution according to the preparation method of the test solution in Example 1. Use the elution gradient programs in Table 1 above and Table 8 below and perform detection under the same other conditions as in Example 1.

[0104] Table 8

[0105] Time (min) Flow rate (ml / min) Mobile phase A (%) Mobile phase B (%) 0-10 1.0 5→10 95→90 10-20 1.0 10 90 20-30 1.0 10→15 90→85 30-56 1.0 15→40 85→60

[0106] The chromatograms obtained by detecting the elution gradient programs in Table 1 and Table 8 are as follows: Fig.12 and Fig.13 The above gradient elution programs can effectively separate characteristic peaks 1-9, but the spectrum obtained by the gradient elution program in Table 1 has richer chromatographic information, better separation of chromatographic peaks, and a more stable baseline, so the gradient elution program in Table 1 is preferred.

[0107] Example 3

[0108] The difference from Example 1 is that the preparation parameters of the test solution are different, as follows:

[0109] 1. Preparation of test solution

[0110] Accurately measure 5 ml of Fuyinkang lotion, place it in a 25 ml volumetric flask, add solvent, adjust to volume, shake well, filter and take the filtrate. Investigate the effect of water and methanol as solvents on the test results, compare the effects of different solvents on the spectrum, and the test results are shown in Fig.14 and Fig.15 shown.

[0111] The results show that by comparing the chromatograms obtained with different solvents, when the solvent is water or methanol, characteristic spectra with stable baselines and more characteristic peak information can be obtained. Among them, the chromatographic peaks obtained with water as the solvent have better peak shape and better economy, so water is selected as the optimal solvent for this experiment.

[0112] Example 4

[0113] The method in Example 1 was tested for precision and stability, and the verification process was as follows:

[0114] 1. Precision inspection

[0115] The same sample solution of Fuyinkang lotion was taken and injected 6 times according to the chromatographic conditions of Example 1 to determine the relative retention time and relative peak area of ​​each common peak. The test results are shown in Table 9.

[0116] Table 9

[0117]

[0118] The results showed that the RSDs of the relative retention times of the characteristic peaks and the reference S peak (peak 3) were all less than 1%, while the RSDs of the relative peak areas were all less than 2%, indicating that the instrumental precision of this method was good.

[0119] 2. Repeatability test

[0120] Take the same portion of Fuyinkang lotion and repeat the preparation of 6 portions according to the test sample preparation method under the chromatographic conditions of Example 1, and measure the relative retention time and relative peak area of ​​each common peak according to the chromatographic conditions. The test results are shown in Table 10.

[0121] Table 10

[0122]

[0123] The results showed that the RSDs of the relative retention times of the characteristic peaks and the reference S peak (peak 3) were all less than 1%, while the RSDs of the relative peak areas were all less than 2%, indicating that the method had good repeatability.

[0124] 3. Stability inspection

[0125] The same Fuyinkang lotion test solution was taken and injected at 0, 3, 6, 12 and 18 hours respectively according to the chromatographic conditions of Example 1 to determine the relative retention time and relative peak area of ​​each common peak. The test results are shown in Table 11.

[0126] Table 11

[0127]

[0128] The results showed that the RSDs of the relative retention times of the characteristic peaks and the reference S peak (peak 3) were all less than 1%, while the RSDs of the relative peak areas were all less than 3%, indicating that the test solution was stable within 18 hours and met the determination requirements.

[0129] Comparative Example 1

[0130] According to the detection method disclosed in the existing document "Research on the Improvement of the Production Process of Fuyinkang Lotion", the specific method is:

[0131] Chromatographic conditions: Inertil ODS-3C 18The chromatographic column was (250 mm × 4.6 mm, 5 μm), the column temperature was 40°C, acetonitrile-0.025 mol / L phosphoric acid solution (8:92) was used as the mobile phase, the detection wavelength was 350 nm, and the theoretical plate number was calculated based on the picrotoxin peak and should not be less than 7000.

[0132] Preparation of reference solution: Accurately weigh an appropriate amount of esculetin control, add methanol to dissolve, and prepare a esculetin standard solution containing 6 μg of esculetin per 1 ml as the reference solution.

[0133] Preparation of test solution: accurately measure 1 ml of the solution and place it in a separatory funnel, add 10 ml of water, shake well, extract with ethyl acetate 4 times, 20 ml each time, combine the ethyl acetate solution, wash with 20 ml of water, and discard the water washing solution. Evaporate the ethyl acetate solution in a water bath, add an appropriate amount of methanol to dissolve the residue, transfer quantitatively to a 10 ml volumetric flask, dilute to the mark with methanol, shake well, accurately measure 1 ml, transfer to a 10 ml volumetric flask, dilute to the mark with methanol, shake well, and obtain.

[0134] Determination method: Accurately aspirate the reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0135] The test results of the test product are as follows Fig.16 As shown, the results show that compared with Example 1, the sample preparation method under the prior art method is cumbersome and the spectral peak is single, which cannot more comprehensively control the quality of Fuyinkang Lotion and has certain limitations.

[0136] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. It should be noted that for those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. A characteristic spectrum detection method for Fuyinkang lotion, characterized in that: The detection was carried out by high performance liquid chromatography, the chromatographic column was a chromatographic column with octadecylsilane bonded silica gel as a filler; the mobile phase was acetonitrile and 0.1% acidic aqueous solution; the elution was gradient elution; The gradient elution is as follows: acetonitrile is used as mobile phase A, 0.1% acidic aqueous solution is used as mobile phase B, and elution is performed according to the following gradient elution program:

2. The characteristic spectrum detection method according to claim 1, characterized in that: The column temperature of the chromatographic column is 29°C-31°C, the flow rate is 0.98mL / min-1.02mL / min; the detection wavelength is 220nm.

3. The characteristic spectrum detection method according to claim 1, characterized in that: The acidic aqueous solution is a phosphoric acid aqueous solution or a formic acid aqueous solution.

4. The characteristic spectrum detection method according to claim 1, characterized in that: The chromatographic column is Kromasil100-5-C18.

5. The characteristic spectrum detection method according to claim 1, characterized in that: The preparation process of the test solution is: take Fuyinkang lotion, add solvent, shake well, and the filtrate obtained after filtering is the test solution.

6. The characteristic spectrum detection method according to claim 5, characterized in that: The solvent is methanol or water.

7. The characteristic spectrum detection method according to claim 1, characterized in that: The preparation process of the reference solution is as follows: take appropriate amounts of matrine, fraxinoyl, fraxinoyl, chlorogenic acid, fraxinoside, and akebia phenylethanol glycoside B reference substances, accurately weigh them, and add methanol to make a mixed reference solution containing 200 μg of matrine, 80 μg of fraxinoyl, 40 μg of fraxinoyl, 40 μg of chlorogenic acid, 60 μg of fraxinoside, and 20 μg of akebia phenylethanol glycoside B per 1 mL.

8. The characteristic spectrum detection method according to claim 1, characterized in that: Taking the chromatographic peak of aesculin as the reference peak, the standard characteristic spectrum of the Fuyinkang lotion comprises 9 peaks, with peak 3 as the S peak, and the retention time of the other characteristic peaks relative to peak 3 is ±10% of the specified value, and the specified value of each characteristic peak is: Peak 1: 0.45, Peak 2: 0.73, Peak 4: 1.30, Peak 5: 1.45, Peak 6: 1.49, Peak 7: 1.84, Peak 8: 1.97, Peak 9: 2.

74.

9. The characteristic spectrum detection method according to claim 1, characterized in that: Taking Peak 3 as the S peak, the relative peak area of ​​Peak 6 shall not be less than 0.

34.

10. The characteristic spectrum detection method according to claim 1, characterized in that: The characteristic spectrum of the Fuyinkang lotion includes characteristic peaks corresponding to matrine, chlorogenic acid, aesculetin, fraxinoside, and akebia phenylethanoid glycoside B; matrine is peak 1, aesculetin is peak 3, chlorogenic acid is peak 4, aesculetin is peak 5, fraxinoside is peak 6, and akebia phenylethanoid glycoside B is peak 8.

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