Quality control method of shuanghuanglian inhalation solution

Abstract

The application relates to the field of traditional Chinese medicine quality control, and particularly discloses a quality control method for Shuanghuanglian inhalation solution. The method can simultaneously determine the fingerprint spectrum and ingredient content of the Shuanghuanglian inhalation solution. The fingerprint spectrum detection method disclosed by the application adopts UPLC determination, has 17 characteristic peaks, and attributes 8 characteristic peaks. The contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C and forsythoside are simultaneously determined. Compared with the prior art, the method for determining the fingerprint spectrum and content of the Shuanghuanglian inhalation solution is improved, and the quality of the Shuanghuanglian inhalation solution can be comprehensively monitored conveniently, quickly and accurately.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical preparations, and more specifically to a quality control method for Shuanghuanglian inhalation solution. Background Technology

[0002] Shuanghuanglian formula is composed of three Chinese herbs: honeysuckle, scutellaria, and forsythia. It is a representative formula for treating respiratory diseases with significant clinical efficacy and has a clinical history of more than 30 years. It has the effects of dispelling wind and releasing the exterior, clearing heat and detoxifying. It is used to treat diseases such as colds, fever, and cough caused by viral and bacterial infections, and has played an important role in the treatment of coronaviruses such as SARS and MERS.

[0003] Shuanghuanglian inhalation solution was developed based on the Shuanghuanglian injection process. It was researched and developed according to the requirements of a single-dose packaged inhaled sterile preparation, and is administered via nebulization. Inhalation solutions for nebulizers refer to solutions in which the drug is dispersed in a suitable medium and delivered to the respiratory tract and lungs in aerosol form via a nebulizer to exert local or systemic therapeutic effects. Inhalation administration significantly reduces the drug concentration entering the systemic bloodstream, thus largely avoiding the serious adverse reactions caused by intravenous administration of injectable drugs.

[0004] Shuanghuanglian inhalation solution is classified as a Class II new traditional Chinese medicine. Currently, there are no published reports in the analytical field regarding the use of UPLC to establish a fingerprint spectrum for Shuanghuanglian inhalation solution and simultaneously determine its component content. Therefore, establishing a rapid, accurate, and comprehensive method for controlling the quality of Shuanghuanglian inhalation solution, and achieving overall control of its material composition, is of great significance. Summary of the Invention

[0005] This invention improves quality control standards from both overall and key component perspectives by using fingerprint spectroscopy for qualitative analysis and quantification of specific active ingredients. Compared to existing technologies, it not only comprehensively and quantitatively determines the controllability of product quality but also significantly shortens the measurement time compared to the commonly used high-performance liquid chromatography (HPLC) technology by employing ultra-high-performance liquid chromatography (UPLC), enabling the complex characterization of the entire product's fingerprint spectroscopy. Furthermore, the combination of UPLC and specific indicative components ensures product quality and qualitative performance while also saving on measurement costs. The test results exhibit excellent reproducibility and stability, are suitable for industrial application, and are ideal for large-scale implementation as a product quality control standard.

[0006] The purpose of this invention is to establish an accurate, sensitive, and reproducible fingerprinting method for Shuanghuanglian inhalation solution as a technical means for quality control. This method can simultaneously determine the content of seven components in Shuanghuanglian inhalation solution, comprehensively characterizing its quality. This allows for better quality control of the preparation and ensures medication safety.

[0007] In view of the problems existing in the prior art, a quality control method for Shuanghuanglian inhalation solution is proposed in this invention.

[0008] To achieve the above objectives, the present invention adopts the following technical solution:

[0009] According to one aspect of the present invention, a quality control method for Shuanghuanglian inhalation solution is provided.

[0010] A quality control method for Shuanghuanglian inhalation solution is disclosed. This method uses UPLC to determine the fingerprint spectrum and content of Shuanghuanglian inhalation solution. The chromatographic conditions of this method are as follows: UPLC C18 column (column length 100 mm, column inner diameter 2.1 mm, particle size 1.8 μm); gradient elution using acetonitrile (A) and 0.3-0.5% phosphoric acid aqueous solution (B) as the mobile phase; flow rate 0.2-0.4 ml / min; injection volume 1-5 μL; column temperature 25-35℃; fingerprint spectrum detection wavelength 200-235 nm; and content determination detection wavelengths 200-235 nm and 300-350 nm.

[0011] The fingerprint pattern detection method further includes the following steps:

[0012] (1) Preparation of reference solution: Accurately weigh an appropriate amount of baicalin reference standard, place it in a volumetric flask, dissolve it in solvent, and dilute to the mark to prepare the reference solution. The solvent includes: 50% methanol, 70% methanol, and methanol.

[0013] (2) Chromatographic conditions: UPLC C18 column (column length 100 mm, column inner diameter 2.1 mm, particle size 1.8 μm); gradient elution with acetonitrile (A) 0.3%–0.4% phosphoric acid aqueous solution (B) as mobile phase; flow rate 0.2–0.4 ml / min; detection wavelength 210 nm; injection volume 1–5 μL; column temperature 25–35 °C.

[0014] Gradient elution: 0-1 min, 7%-10% A; 1-3.5 min, 10%-11% A; 3.5-5 min, 11%-14% A; 5-10 min, 14%-15% A; 10-16 min, 15%-17% A; 16-19 min, 17%-25% A; 19-30 min, 25% A.

[0015] (3) Establishment of standard fingerprint spectrum: By measuring the UPLC fingerprint spectrum of 10 batches of Shuanghuanglian inhalation solution, the common fingerprint characteristics were determined, and the standard fingerprint spectrum of Shuanghuanglian inhalation solution was established by fitting with the Chinese medicine fingerprint spectrum similarity evaluation software.

[0016] (4) Quality control of fingerprint chromatograms: The UPLC fingerprint chromatograms of the samples are compared with standard fingerprint chromatograms. The similarity calculated by the traditional Chinese medicine fingerprint chromatogram similarity evaluation software should not be less than 0.9.

[0017] The fingerprint spectrum was detected by UPLC and showed 17 characteristic peaks. The relative retention times of baicalin were as follows: 0.136, 0.147, 0.220, 0.235, 0.251, 0.270, 0.541, 0.647, 0.672, 0.737, 0.801, 0.836, 0.934, 1.000, 1.028, 1.134, and 1.143.

[0018] According to another aspect of the present invention, a method for determining the content of an inhaled solution is provided, comprising the following steps:

[0019] (1) Preparation of reference solutions: Accurately weigh appropriate amounts of several or all of the reference standards for neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, and forsythoside, place them in a volumetric flask, dissolve them in 50% methanol, and dilute to the mark to prepare the reference solution stock solution. Accurately pipette the reference solution stock solution into a volumetric flask and dilute to the mark with 50% methanol to prepare a mixed reference solution.

[0020] (2) Preparation of test solution: Accurately pipette 1-5 ml of Shuanghuanglian inhalation solution into a 50 ml volumetric flask, add 50% methanol to the mark, shake well, filter through a 0.22 μm microporous membrane, and take the filtrate as the test solution.

[0021] (3) Chromatographic conditions: UPLC C18 column (column length 100 mm, column inner diameter 2.1 mm, particle size 1.8 μm); gradient elution with acetonitrile (A)-0.4% phosphoric acid aqueous solution (B) as mobile phase; flow rate 0.2-0.4 ml / min; injection volume 1-5 μL; column temperature 25-35℃; PDA detector; detection wavelength 210 nm, 324 nm.

[0022] (4) Determination method: Take 1-5 μl of the reference solution and the test solution respectively, inject them into the liquid chromatograph, record the chromatogram, and the result is obtained.

[0023] The chromatographic conditions for step (3) include the following detection wavelengths: 210 nm for forsythoside, and 324 nm for neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, and isochlorogenic acid C.

[0024] The beneficial effects of this invention are:

[0025] This invention establishes a detection method for the fingerprint spectrum of Shuanghuanglian inhalation solution, identifying 17 common fingerprint peaks for ultraviolet detection, which fully characterizes the integrity and complexity of traditional Chinese medicine preparations.

[0026] The method of the present invention can simultaneously recover seven components in Shuanghuanglian inhalation solution: neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, and forsythoside, with an average recovery rate of 93.9% to 103.4% and an RSD of less than 5.0%.

[0027] When using acetonitrile-0.4% phosphoric acid solution as the mobile phase, the baseline is relatively stable, the chromatographic peaks are symmetrical, and the resolution is good. The resolution of each component in the content determination is greater than 1.5.

[0028] While ensuring accuracy and effectiveness, it also takes into account the principle of cost saving, making it more suitable for large-scale production and improving the existing quality control standards for Shuanghuanglian inhalation solution. Attached Figure Description

[0029] Figure 1 This is a reference fingerprint of Shuanghuanglian inhalation solution;

[0030] Figure 2 The results of the fingerprint spectrum similarity study of different batches of Shuanghuanglian inhalation solution;

[0031] Figure 3 Peak assignment chromatogram of Shuanghuanglian inhalation solution;

[0032] Figure 4 Peak identification spectrum for Shuanghuanglian inhalation solution;

[0033] Figure 5 Reference material and 235nm fingerprint spectrum of Shuanghuanglian inhalation solution;

[0034] Figure 6 Reference material and 200nm fingerprint spectrum of Shuanghuanglian inhalation solution. Detailed Implementation

[0035] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be described clearly and completely below in conjunction with specific embodiments.

[0036] The Shuanghuanglian inhalation solution used in the following examples is a product of Yingkerui (Hengqin) Pharmaceutical Research Institute Co., Ltd.

[0037] According to application number CN201510854860.6, invention title: A solution preparation for Shuanghuanglian nebulized inhalation and its preparation method, it is prepared.

[0038] Example 1: Establishment of fingerprint spectrum for Shuanghuanglian inhalation solution

[0039] Instruments: Liquid Chromatograph: Agilent 1290 Infinity II; Electronic Analytical Balance: Sartorius Scientific Instruments (Beijing) Co., Ltd., SQP (1 / 100,000)

[0040] Test drug: Shuanghuanglian inhalation solution (batch numbers: CP-200527-02, CP-200715-01, CP-200716-01, CP-200717-01, CP-201025-01, CP-201101-01, CP-201102-01, CP-201210-01, CP-201210-02, CP-201210-03), produced by Yingkerui (Hengqin) Pharmaceutical Research Institute.

[0041] Baicalin reference standard (batch number: 110715-201721, purity: 95.4%) was purchased from the National Institutes for Food and Drug Control.

[0042] New chlorogenic acid reference standard (batch number: DST200521-015, purity (self-tested): 97.6%), purchased from Chengdu Lemeitian Pharmaceutical | Desite Biotechnology Co., Ltd.

[0043] Chlorogenic acid reference standard (batch number: 110753-201817, purity: 96.8%) was purchased from the National Institutes for Food and Drug Control.

[0044] Cryptochlorogenic acid reference standard (batch number: DST200521-035, purity (self-tested): 97.5%) was purchased from Chengdu Lemeitian Pharmaceutical | Desite Biotechnology Co., Ltd.

[0045] Isochlorogenic acid B reference standard (batch number: 000319-201912, purity (self-tested): 98.3%) was purchased from Jiangxi Baicaoyuan Biotechnology Co., Ltd.

[0046] Isochlorogenic acid A reference standard (batch number: 111782-201807, purity: 94.3%) was purchased from the National Institutes for Food and Drug Control.

[0047] Isochlorogenic acid C reference standard (batch number: 111894-201102, purity: 94.1%) was purchased from the National Institutes for Food and Drug Control.

[0048] Forsythoside reference standard (batch number: 110821-201816, purity: 95.1%) was purchased from the National Institutes for Food and Drug Control.

[0049] Acetonitrile was chromatographically pure and manufactured by OCEANPAK; phosphoric acid was chromatographically pure and manufactured by Tianjin Kemei Chemical Reagent Co., Ltd.; methanol was analytically pure and manufactured by Guangzhou Chemical Reagent Factory; and water was ultrapure water manufactured by Shanghai Hetai Instrument Co., Ltd.

[0050] The fingerprint chromatogram was detected using ultra-high performance liquid chromatography (UHPLC) under the following specific conditions:

[0051] Chromatographic conditions: Octadecylsilane-bonded silica gel was used as the stationary phase; UHPLC XB C18 column (column length 100 mm, column inner diameter 2.1 mm, particle size 1.8 μm); gradient elution was performed using acetonitrile (A)-0.4% phosphoric acid aqueous solution (B) as the mobile phase, with a detection wavelength of 210 nm; column temperature 30 ℃; flow rate 0.3 ml per minute.

[0052] Gradient elution: 0-1 min, 7%-10% A; 1-3.5 min, 10%-11% A; 3.5-5 min, 11%-14% A; 5-10 min, 14%-15% A; 10-16 min, 15%-17% A; 16-19 min, 17%-25% A; 19-30 min, 25% A.

[0053] Preparation of reference solution: Take an appropriate amount of baicalin reference standard, accurately weigh it, add methanol to prepare a solution containing 0.2 mg per ml, shake well, and the solution is ready.

[0054] Preparation of the test solution: Accurately measure 1 ml of this product, place it in a 50 ml volumetric flask, dilute to the mark with 50% methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0055] Determination method: Take 2 μl of the reference solution and the test solution respectively, inject them into the liquid chromatograph, and determine the result.

[0056] Establishment of fingerprint profiles

[0057] Ten batches of Shuanghuanglian inhalation solution were prepared according to the preparation method of Shuanghuanglian inhalation solution. Ten batches of test solution were also prepared using the same method. The fingerprint chromatograms of each test solution were recorded by injecting them into a liquid chromatogram. These fingerprint chromatograms were then imported into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system to generate a control fingerprint chromatogram. The similarity between the ten batches of Shuanghuanglian inhalation solution and the control fingerprint chromatogram was calculated based on common peaks. The results are shown in [Figure Number]. Figure 1 , Figure 2 Table 1.

[0058] Table 1. Results of fingerprint spectrum similarity analysis of different batches of Shuanghuanglian inhalation solution

[0059] R S2 S3 S4 S5 S6 S7 S8 S9 S10 S11 R 1.000 0.993 0.995 0.994 0.994 0.997 0.997 0.997 0.998 0.994 0.994 S2 0.993 1.000 0.991 0.989 0.989 0.984 0.984 0.984 0.988 0.984 0.987 S3 0.995 0.991 1.000 1.000 0.999 0.986 0.986 0.986 0.989 0.978 0.979 S4 0.994 0.989 1.000 1.000 0.999 0.985 0.985 0.985 0.988 0.976 0.977 S5 0.994 0.989 0.999 0.999 1.000 0.986 0.986 0.986 0.988 0.979 0.979 S6 0.997 0.984 0.986 0.985 0.986 1.000 1.000 1.000 0.988 0.996 0.995 S7 0.997 0.984 0.986 0.985 0.986 1.000 1.000 1.000 0.988 0.996 0.995 S8 0.997 0.984 0.986 0.985 0.986 1.000 1.000 1.000 0.998 0.996 0.995 S9 0.998 0.988 0.989 0.988 0.988 0.998 0.998 0.998 1.000 0.996 0.997 S10 0.994 0.984 0.978 0.976 0.979 0.996 0.996 0.996 0.996 1.000 0.999 S11 0.994 0.987 0.979 0.977 0.979 0.995 0.995 0.995 0.997 0.999 1.000

[0060] R: Compare fingerprint chromatograms, S2: CP-200527-02, S3: CP-200715-01, S4: CP-200716-01, S5: CP-200717-01, S6: CP-201025-01, S7: CP-201101-01, S8: CP-201102-01, S9: CP-201210-01, S10: CP-201210-02, S11: CP-201210-03

[0061] Based on the similarity calculated from the fingerprint chromatograms, the similarity between the fingerprint chromatograms of 10 batches of Shuanghuanglian inhalation solution and the reference fingerprint chromatograms ranged from 0.993 to 0.998, with all similarities greater than 0.90. The fingerprint chromatogram of the test sample should show chromatographic peaks with the same retention time as the reference chromatographic peaks. The chromatogram of the test sample should be basically consistent with the reference fingerprint chromatogram, with 17 corresponding common peaks. According to the similarity evaluation system for chromatographic fingerprint chromatograms of traditional Chinese medicine, the similarity is calculated based on the common peaks. The similarity between the fingerprint chromatogram of the test sample and the reference fingerprint chromatogram should not be less than 0.90.

[0062] Peak identification and assignment based on fingerprint spectrum

[0063] Following the fingerprint chromatographic determination method, the following solutions were injected separately: Shuanghuanglian inhalation solution (test sample), honeysuckle and forsythia suspensa single-herb decoction solution, scutellaria baicalensis extract solution, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, forsythoside mixed reference solution, and baicalin reference solution. Chromatograms were recorded to determine the assignment of each chromatographic peak in the Shuanghuanglian inhalation solution. Experimental results are shown below. Figure 3 , Figure 4 Table 2.

[0064] Table 2. Attribution of chromatographic peaks in fingerprint chromatograms

[0065]

[0066] Example 2: Determination of the content of seven components in Shuanghuanglian inhalation solution

[0067] The ultra-high performance liquid chromatography-dual-wavelength detection method was used, with the following specific detection conditions:

[0068] Chromatographic conditions: Octadecylsilane-bonded silica gel was used as the stationary phase; UHPLC XB C18 column (column length 100 mm, inner diameter 2.1 mm, particle size 1.8 μm); acetonitrile (A) - 0.4 Gradient elution was performed using a % phosphoric acid aqueous solution (B) as the mobile phase. Neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, and isochlorogenic acid C were detected at a wavelength of 324 nm, and forsythoside was detected at a wavelength of 210 nm. The column temperature was 30 °C, and the flow rate was 0.3 ml per minute.

[0069] Gradient elution: Same as in Example 1.

[0070] Preparation of reference solution: Take appropriate amounts of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, and forsythoside reference standards, accurately weigh them, add 50% methanol to prepare a mixed reference solution containing 15 μg of neochlorogenic acid, 12 μg of chlorogenic acid, 15 μg of cryptochlorogenic acid, 6 μg of isochlorogenic acid B, 3 μg of isochlorogenic acid A, 6 μg of isochlorogenic acid C, and 15 μg of forsythoside per 1 ml, shake well, and the solution is ready.

[0071] Preparation of the test solution: Accurately measure 1 ml of this product, place it in a 50 ml volumetric flask, dilute to the mark with 50% methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0072] Determination method: Inject 2 μl of the reference solution and the test solution into the liquid chromatograph, respectively, and determine the result. Experimental results are shown in Table 3.

[0073] Table 3. Content determination results

[0074]

[0075] Example 3, Fingerprint determination of Shuanghuanglian inhalation solution 1

[0076] The ultra-high performance liquid chromatography-dual-wavelength detection method was used, with the following specific detection conditions:

[0077] Chromatographic conditions: Octadecylsilane-bonded silica gel was used as the stationary phase; UHPLC XB C18 column (column length 100 mm, inner diameter 2.1 mm, particle size 1.8 μm); acetonitrile (A) - 0.3 Gradient elution was performed using a % phosphoric acid aqueous solution (B) as the mobile phase, with a detection wavelength of 235 nm, a column temperature of 30 °C, and a flow rate of 0.3 ml per minute.

[0078] Gradient elution: Same as the content under "Gradient elution" in Example 1.

[0079] Preparation of reference solution: Same as the content under "Preparation of reference solution" in Example 1.

[0080] Preparation of test solution: Accurately measure 1 ml of this product, place it in a 50 ml volumetric flask, dilute with methanol to the mark, shake well, filter, and take the filtrate to obtain the test solution.

[0081] Determination method: Inject 2 μl of the reference solution and the test solution into the liquid chromatograph, respectively, and determine the result. See the experimental results below. Figure 5 .

[0082] Example 4, Determination of the content of Shuanghuanglian inhalation solution 1

[0083] The ultra-high performance liquid chromatography-dual-wavelength detection method was used, with the following specific detection conditions:

[0084] Chromatographic conditions: Octadecylsilane-bonded silica gel was used as the stationary phase; UHPLC XB C18 column (column length 100 mm, column inner diameter 2.1 mm, particle size 1.8 μm); gradient elution was performed using acetonitrile (A)-0.3% phosphoric acid aqueous solution (B) as the mobile phase; the detection wavelengths for neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, and isochlorogenic acid C were 350 nm, and the detection wavelength for forsythoside was 235 nm; the column temperature was 30 ℃; and the flow rate was 0.3 ml per minute.

[0085] Gradient elution procedure: Same as in Example 1.

[0086] Preparation of reference solution: Same as the content under "Preparation of reference solution" in Implementation Case 2.

[0087] Preparation of test solution: Accurately measure 1 ml of this product, place it in a 50 ml volumetric flask, dilute with methanol to the mark, shake well, filter, and take the filtrate to obtain the test solution.

[0088] Determination method: Inject 2 μl of the reference solution and the test solution into the liquid chromatograph, respectively, and determine the result. Experimental results are shown in Table 4.

[0089] Table 4. Content determination results

[0090]

[0091]

[0092] Example 5, Fingerprint determination of Shuanghuanglian inhalation solution 2

[0093] The ultra-high performance liquid chromatography-dual-wavelength detection method was used, with the following specific detection conditions:

[0094] Chromatographic conditions: Octadecylsilane-bonded silica gel was used as the stationary phase; UHPLC XB C18 column (column length 100 mm, column inner diameter 2.1 mm, particle size 1.8 μm); gradient elution was performed using acetonitrile (A)-0.5% phosphoric acid aqueous solution (B) as the mobile phase, with a detection wavelength of 200 nm; column temperature 30 ℃; flow rate 0.3 ml per minute.

[0095] Gradient elution: Same as in Example 1.

[0096] Preparation of reference solution: Same as the content under "Preparation of reference solution" in Example 1.

[0097] Preparation of test solution: Accurately measure 1 ml of this product, place it in a 50 ml volumetric flask, dilute to the mark with 70% methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0098] Determination method: Inject 2 μl of the reference solution and the test solution into the liquid chromatograph, respectively, and determine the result. See the experimental results below. Figure 6 .

[0099] Example 6, Determination of the content of Shuanghuanglian inhalation solution 2

[0100] The ultra-high performance liquid chromatography-dual-wavelength detection method was used, with the following specific detection conditions:

[0101] Chromatographic conditions: Octadecylsilane-bonded silica gel was used as the stationary phase; UHPLC XB C18 column (column length 100 mm, column inner diameter 2.1 mm, particle size 1.8 μm); gradient elution was performed using acetonitrile (A)-0.5% phosphoric acid aqueous solution (B) as the mobile phase; the detection wavelengths for neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, and isochlorogenic acid C were 300 nm, and the detection wavelength for forsythoside was 200 nm; the column temperature was 30 ℃; and the flow rate was 0.3 ml per minute.

[0102] Gradient elution: Same as in Example 1.

[0103] Preparation of reference solution: Same as the content under "Preparation of reference solution" in Example 2.

[0104] Preparation of test solution: Accurately measure 1 ml of this product, place it in a 50 ml volumetric flask, dilute with methanol to the mark, shake well, filter, and take the filtrate to obtain the test solution.

[0105] Determination method: Inject 2 μl of the reference solution and the test solution into the liquid chromatograph, respectively, and determine the result. Experimental results are shown in Table 5.

[0106] Table 5. Content determination results

[0107]

[0108] The present invention has been described in detail above. The description of the embodiments above is only for the purpose of helping to understand the method and core idea of ​​the present invention. It should be noted that those skilled in the art can make several improvements and modifications to the present invention without departing from the principle of the present invention, and these improvements and modifications also fall within the protection scope of the claims of the present invention.

Claims

1. A quality control method for Shuanghuanglian inhalation solution, characterized in that, This method uses UPLC to determine the fingerprint spectrum and content of Shuanghuanglian inhalation solution. The chromatographic conditions of this detection method are: UPLC C18 column, column length 100 mm, column inner diameter 2.1 mm, particle size 1.8 μm; Acetonitrile (A) and 0.4% phosphoric acid aqueous solution (B) were used as the mobile phase for gradient elution. The gradient elution was as follows: 0–1 min, 7%–10% A; 1–3.5 min, 10%–11% A; 3.5–5 min, 11%–14% A; 5–10 min, 14%–15% A; 10–16 min, 15%–17% A; 16–19 min, 17%–25% A; 19–30 min, 25% A. The flow rate was 0.2–0.4 ml / min. The injection volume was 1–5 μL. The column temperature was 25–35 °C. The fingerprint detection wavelength was 210 nm, and the content determination detection wavelengths were 210 nm and 324 nm.

2. The quality control method for Shuanghuanglian inhalation solution according to claim 1, characterized in that, The fingerprint pattern detection method further includes the following steps: (1) Preparation of reference solution: Weigh an appropriate amount of baicalin reference standard accurately, place it in a volumetric flask, add solvent to dissolve it and dilute to the mark to prepare a reference solution; (2) Chromatographic conditions: UPLC C18 column, column length 100 mm, column inner diameter 2.1 mm, particle size 1.8 μm; gradient elution with acetonitrile (A) and 0.4% phosphoric acid aqueous solution (B) as mobile phase; flow rate 0.2–0.4 ml / min; detection wavelength 210 nm; injection volume 1–5 μL; column temperature 25–35 °C; Gradient elution: 0–1 min, 7%–10% A; 1–3.5 min, 10%–11% A; 3.5–5 min, 11%–14% A; 5–10 min, 14%–15% A; 10–16 min, 15%–17% A; 16–19 min, 17%–25% A; 19–30 min, 25% A; (3) Establishment of standard fingerprint spectrum: By measuring the UPLC fingerprint spectrum of 10 batches of Shuanghuanglian inhalation solution, the common fingerprint characteristics were determined, and the standard fingerprint spectrum of Shuanghuanglian inhalation solution was established by fitting with the Chinese medicine fingerprint spectrum similarity evaluation software. (4) Quality control of fingerprint spectrum: The UPLC fingerprint spectrum of the sample is compared with the standard fingerprint spectrum. The similarity calculated by the Chinese medicine fingerprint spectrum similarity evaluation software should not be less than 0.

9.

3. The quality control method for Shuanghuanglian inhalation solution according to claim 2, characterized in that, The fingerprint spectrum detection, performed using UPLC, showed 17 characteristic peaks. The relative retention times of these characteristic peaks for baicalin were as follows: 0.136, 0.147, 0.220, 0.235, 0.251, 0.270, 0.541, 0.647, 0.672, 0.737, 0.801, 0.836, 0.934, 1.000, 1.028, 1.134, and 1.

143.

4. The quality control method for Shuanghuanglian inhalation solution according to claim 1, characterized in that, The content determination method comprises the following steps: (1) Preparation of reference solution: Accurately weigh appropriate amounts of several or all of the reference standards of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, and forsythoside, place them in a volumetric flask, add 50% methanol to dissolve and dilute to the mark to prepare the reference solution stock solution; accurately pipette the reference solution stock solution, place it in a volumetric flask and add 50% methanol to dilute to the mark to prepare a mixed reference solution; (2) Preparation of test solution: Accurately pipette 1-5 ml of Shuanghuanglian inhalation solution into a 50 ml volumetric flask, add 50% methanol to the mark, shake well, filter through a 0.22 μm microporous membrane, and take the filtrate as the test solution. (3) Chromatographic conditions: UPLC C18 column, column length 100 mm, column inner diameter 2.1 mm, particle size 1.8 μm; gradient elution with acetonitrile (A) and 0.4% phosphoric acid aqueous solution (B) as mobile phase; flow rate 0.2-0.4 ml / min; injection volume 1-5 μL; column temperature 25-35℃; PDA detector, detection wavelength 210 nm, 324 nm; (4) Determination method: Take 1-5 μl of the reference solution and the test solution respectively, inject them into the liquid chromatograph, record the chromatogram, and the result is obtained.

5. The method for determining the content of Shuanghuanglian inhalation solution according to claim 4, characterized in that: The chromatographic conditions for step (3) include the following detection wavelengths: 210 nm for forsythoside, and 324 nm for neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, and isochlorogenic acid C.

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