UPLC analysis method for related substances of fluticasone propionate inhalation aerosol

Abstract

The invention belongs to the technical field of drug detection, and relates to a UPLC analysis method for related substances of fluticasone propionate inhalation aerosol. Comprising the following steps: mixing acetonitrile and phosphoric acid water to prepare a diluent, adopting the diluent to dissolve fluticasone propionate inhalation aerosol to prepare a sample solution, adopting a chromatographic column taking octadecylsilane chemically bonded silica as a filling agent, taking the phosphoric acid water as a mobile phase A and taking acetonitrile as a mobile phase B, and carrying out gradient elution to obtain the fluticasone propionate inhalation aerosol. And performing UPLC detection on the sample solution according to a gradient elution program. Through the method, 12 impurities in the fluticasone propionate inhalation aerosol can be completely separated and accurately determined. The method is good in specificity and accurate in quantification, meanwhile, the mobile phase is simple and efficient to prepare, detection can be completed only in 13 min, the analysis time is greatly shortened, the solvent dosage is reduced, and the analysis cost is reduced.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of drug detection, and relates to an UPLC (ultra-performance liquid chromatography) analysis method for related substances of fluticasone propionate inhalation aerosol. BACKGROUND

[0002] The information disclosed in this Background section is only for the purpose of increasing the understanding of the general background of the application and does not necessarily constitute an admission that the information forms part of the prior art already known to a person of ordinary skill in the art.

[0003] Fluticasone propionate aerosol is an inhaled glucocorticoid drug, mainly used for prophylactic treatment of bronchial asthma, controlling airway inflammation through anti-inflammatory effect, and reducing the frequency of asthma attacks. Compared with other dosage forms of fluticasone propionate (such as nasal spray, aerosol suspension, dry powder inhalation, etc.), fluticasone propionate aerosol has obvious advantages in lung drug deposition efficiency, onset speed, dose flexibility and portability, and is especially suitable for asthma patients who need long-term maintenance treatment.

[0004] However, according to the research and understanding of the inventors, the quality standard of fluticasone propionate aerosol is not included in the 2025 edition of Chinese Pharmacopoeia, and the import registration standard JX20190073 and USP Pharmacopoeia are included and the related substance detection methods are consistent, which has the following problems: the mobile phase is A, B and C three phases, the detection time is as long as 70 minutes and does not return to the gradient, the running time is relatively long, and the analysis cost is relatively high; the separation degree of impurities GR40775X and impurities CCI18773 is poor, and the chromatographic peaks are basically coincident. Therefore, it is necessary to establish a set of specific, reproducible, accurate and simple and fast related substance detection and analysis method, so as to control the quality of fluticasone propionate inhalation aerosol related substances. SUMMARY

[0005] In order to solve the problems of the related substance detection of fluticasone propionate aerosol, the purpose of the present application is to provide an UPLC analysis method for related substances of fluticasone propionate inhalation aerosol. The UPLC analysis method of the present application has the advantages of simple mobile phase preparation, short analysis time, less solvent consumption, low analysis cost, high separation degree between impurities, etc., and can meet the requirements of drug detection accuracy.

[0006] In order to achieve the above-mentioned purpose, the technical scheme of the present application is as follows: In a first aspect, an UPLC analysis method for related substances of fluticasone propionate inhalation aerosol is provided, comprising the following steps: The acetonitrile and the 0.030-0.070% phosphoric acid water with a volume percentage are mixed in a proportion of 1:0.9-1.1 to prepare a diluent, and the fluticasone propionate inhalation aerosol is dissolved in the diluent to prepare a sample solution, The chromatographic column filled with octadecylsilane bonded silica gel is used as a filler, the 0.030-0.070% phosphoric acid water with a volume percentage is used as mobile phase A, and the acetonitrile is used as mobile phase B, and the sample solution is subjected to UPLC detection according to the gradient elution program in the following table;

[0007] The diluent prepared by mixing the acetonitrile and the 0.030-0.070% phosphoric acid water with a volume percentage in a proportion of 1:0.9-1.1 can not only dissolve the fluticasone propionate inhalation aerosol, but also reduce the matrix effect in the UPLC detection process.

[0008] The 0.030-0.070% phosphoric acid water with a volume percentage is used as mobile phase A, and the acetonitrile is used as mobile phase B according to the specific gradient elution program, so that the raw material and each impurity can be quickly separated, and the UPLC detection of the related substances of the fluticasone propionate inhalation aerosol can be realized.

[0009] In the second aspect, a UPLC detection kit for the related substances of the fluticasone propionate inhalation aerosol comprises: The diluent is prepared by mixing the acetonitrile and the 0.030-0.070% phosphoric acid water with a volume percentage in a proportion of 1:0.9-1.1; The mobile phase is that the 0.030-0.070% phosphoric acid water with a volume percentage is used as mobile phase A, and the acetonitrile is used as mobile phase B; The carrier is used to record the UPLC detection conditions, and the UPLC detection conditions include that the chromatographic column filler is octadecylsilane bonded silica gel, and the gradient elution program is shown in the following table:

[0010] In the third aspect, the UPLC analysis method or the UPLC detection kit is applied to the quality monitoring of the related substances of the fluticasone propionate inhalation aerosol in the production process of the fluticasone propionate inhalation aerosol or in the storage process of the fluticasone propionate inhalation aerosol.

[0011] The beneficial effects of the present application are: 1. Compared with the prior art method for detecting related substances of fluticasone propionate inhalation aerosol, the method of the present application adopts only a mobile phase of 0.040-0.060% (volume fraction) phosphoric acid water and acetonitrile, the preparation process is simple, the preparation time of the mobile phase is saved, the analysis time is shortened from 80 min to 13 min, the analysis time is greatly shortened, the solvent consumption is reduced, the analysis cost is reduced, and the requirements of drug detection accuracy can be met.

[0012] 2. The analysis method of the present application can effectively detect 12 known impurities in fluticasone propionate inhalation aerosol, and the separation effect between each impurity and between the main component and the adjacent impurity is good, solving the problem of preparing two different system suitability solutions due to the low separation degree of impurity GR40775X and impurity CCI18773 in the original method, and solving the problem of low separation degree of unknown impurities and known impurity GR68620X in sample high temperature degradation, ensuring accurate impurity positioning and accurate calculation during sample detection.

[0013] 3. When the fluticasone propionate inhalation aerosol is detected by the method of the present application, the method has good specificity, and the blank excipient does not interfere with the detection of the content of the main component and impurities. BRIEF DESCRIPTION OF DRAWINGS

[0014] The drawings accompanying the specification of the present application serve to provide a further understanding of the present application, and the illustrative embodiments of the present application and their descriptions serve to explain the present application, and do not constitute an improper limitation of the present application.

[0015] Figure 1 System suitability solution spectrum of the comparative example of the present application; Figure 2 Impurity positioning comparison spectrum of the comparative example of the present application; Figure 3 System suitability positioning spectrum of the embodiment of the present application; Figure 4 Specificity overlay of the embodiment of the present application. DETAILED DESCRIPTION

[0016] It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as generally understood by those skilled in the art to which the present application belongs.

[0017] It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments in accordance with the present application. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms "comprises" and / or "comprising," when used in this specification, specify the presence of stated features, steps, operations, elements, components, and / or groups thereof, but do not preclude the presence or addition of one or more other features, steps, operations, elements, components, and / or groups thereof.

[0018] The phosphoric acid water with a volume percentage concentration of 0.030-0.070% in the present application refers to a volume of 0.3-0.7ml of a phosphoric acid solution (with a mass percentage concentration of 85%) in 1000ml of solution.

[0019] The impurity structure and impurity code of fluticasone propionate in the present application are shown in the following table:

[0020] In view of the problems such as long analysis time and poor impurity separation degree of the existing fluticasone propionate aerosol related substance detection method, the present application provides a UPLC analysis method for fluticasone propionate inhalation aerosol related substances.

[0021] In a typical embodiment of the present application, a UPLC analysis method for fluticasone propionate inhalation aerosol related substances is provided, which comprises the following steps: Acetonitrile and phosphoric acid water with a volume percentage concentration of 0.030-0.070% are mixed in a volume ratio of 1:0.9-1.1 to prepare a diluent, and the diluent is used to dissolve fluticasone propionate inhalation aerosol to prepare a sample solution, An octadecylsilane-bonded silica gel chromatographic column is used as a filler, and phosphoric acid water with a volume percentage concentration of 0.030-0.070% is used as mobile phase A, and acetonitrile is used as mobile phase B, and the sample solution is subjected to UPLC detection according to the gradient elution program in the following table;

[0022] In some embodiments, the substances detected for analysis include fluticasone propionate and fluticasone propionate aerosol impurities. The fluticasone propionate aerosol impurities are: 、 、 、 、 、 、 、 、 、 、 、 .

[0023] In some embodiments, the gradient elution procedure is shown in the following table:

[0024] In some embodiments, the chromatographic column used in UPLC detection is ACQUITY UPLC BEH C18, 2.1mm x 100mm, 1.7μm, or an equivalent performance chromatographic column.

[0025] In some embodiments, the detection wavelength in UPLC detection is 229-249nm, preferably 239nm.

[0026] In some embodiments, the flow rate in UPLC detection is 0.50-0.70ml / min, preferably 0.60ml / min.

[0027] In some embodiments, the column temperature of the chromatographic column in UPLC detection is 40-50℃, preferably 45℃.

[0028] In some embodiments, the injection volume in UPLC detection is 3-5μl, preferably 4μl.

[0029] The second embodiment of the present application provides a UPLC detection kit for related substances of fluticasone propionate inhalation aerosol, comprising: The diluent is prepared by mixing acetonitrile and 0.030-0.070% (v / v) phosphoric acid water in a ratio of 1:0.9-1.1; The mobile phase is 0.030-0.070% (v / v) phosphoric acid water as mobile phase A and acetonitrile as mobile phase B; The carrier is used to record the conditions of UPLC detection; the conditions of UPLC detection include that the chromatographic column filler is octadecylsilane bonded silica gel, and the gradient elution procedure; the gradient elution procedure is shown in the following table:

[0030] In some embodiments, it further comprises impurity control.

[0031] The third embodiment of the present application provides the application of the above-mentioned UPLC analysis method or UPLC detection kit in the quality monitoring of related substances of fluticasone propionate inhalation aerosol during the production of fluticasone propionate inhalation aerosol or during the storage of fluticasone propionate inhalation aerosol.

[0032] In order to enable those skilled in the art to more clearly understand the technical solutions of the present application, the technical solutions of the present application will be described in detail below in combination with specific examples and comparative examples.

[0033] Reference: Standard test method (HPLC) 1-1 Chromatographic conditions Mobile phase A: acetonitrile containing 0.05% phosphoric acid (0.5ml of a 85% phosphoric acid solution in 1000ml of acetonitrile); Mobile phase B: methanol containing 0.05% phosphoric acid (0.5ml of a 85% phosphoric acid solution in 1000ml of methanol); Mobile phase C: 0.05% phosphoric acid water (0.5ml of a 85% phosphoric acid solution in 1000ml of water); Diluent: mobile phase A-mobile phase C (1:1); Chromatographic column: Inertsil ODS-24.6*250mmmm, 5μm; Detection wavelength: 239nm; Flow rate: 1.0ml / min; Column temperature: 40℃; Injection volume: 20μL.

[0034] Elution mode: gradient elution;

[0035] 1-2 Preparation of solutions System suitability solution: one fluticasone propionate sample placed at high temperature 50℃ for 30 days was dissolved with diluent, and appropriate amounts of impurity reference substances were added to prepare a mixed solution containing 0.22mg of fluticasone propionate and 0.4μg of other impurities per 1ml.

[0036] Impurity GR40775X positioning solution: appropriate amount of impurity GR40775X reference substance was dissolved with diluent to prepare a 0.1μg / ml solution.

[0037] Impurity CCI18773 positioning solution: appropriate amount of CCI18773 reference substance was dissolved with diluent to prepare a 0.1μg / ml solution.

[0038] 1-3 Experimental conclusion: The results of the system suitability solution are shown in Table 1, and the chromatogram is shown in Figure 1 and Figure 2 . Impurity GR40775X and impurity CCI18773 completely coincide, and the high-temperature degradation impurity basically coincides with known impurity GR68620. The separation degree cannot be calculated, and the problem of impurity positioning and accurate calculation is very easy to cause during the actual sample detection process. In addition, the running time is too long, and the gradient regression time needs 80min, which is too high in analysis cost.

[0039] Table 1 System Suitability Solution Results

[0040] Note: "-" means not applicable, same below.

[0041] Example 1: Method of the Invention (UPLC) 1-1 Chromatographic Conditions Mobile Phase A: 0.05% phosphoric acid water; Mobile Phase B: acetonitrile.

[0042] Chromatographic Column: ACQUITY UPLC BEH C18, 2.1 mm x 100 mm, 1.7 μm.

[0043] Elution Mode: Gradient elution;

[0044] Detection wavelength was 239 nm; Flow rate was 0.60 ml / min; Column temperature was 45 °C; Injection volume was 4 μL.

[0045] 1-2 Solution Preparation Diluent: Mobile Phase A and Mobile Phase B were mixed at a volume ratio of 1:1.

[0046] System Suitability Solution: One fluticasone propionate sample that was placed at high temperature 50 °C for 30 days was dissolved with diluent, and appropriate amounts of impurity reference substances were added to prepare a mixed solution containing 0.22 mg of fluticasone propionate and 0.4 μg of each of the other impurities per 1 ml.

[0047] Impurity GR40775X Positioning Solution: Appropriate amounts of impurity GR40775X reference substance were dissolved with diluent to prepare a 0.1 μg / ml solution.

[0048] Impurity CCI18773 Positioning Solution: Appropriate amounts of CCI18773 reference substance were dissolved with diluent to prepare a 0.1 μg / ml solution.

[0049] Blank Excipient Solution (labeled as 20250427-6-0d): One self-prepared fluticasone propionate blank excipient was opened after being deflated, 40 ml of diluent was added, and it was shaken to obtain the solution.

[0050] 1-3 Experimental Conclusion: System Suitability Solution Results are shown in Table 2, and the chromatogram is shown in Figure 3 and Figure 4The blank adjuvant solution does not interfere with the separation and detection of fluticasone propionate and each impurity; the separation degree of the difficult-to-separate impurity GR40775X and the impurity CCI18773 in the system suitability solution is 1.63, the separation degree of the unknown impurity and the known impurity GR68620X in the high-temperature degradation is 1.57, which meets the separation requirement of impurities, can accurately locate and calculate each impurity, and the running time is shortened from 80 min to 13 min, greatly saving the analysis cost.

[0051] Table 2 System suitability

[0052] In combination with the above experimental results, the UPLC detection method of the present application is used for detecting fluticasone propionate, the mobile phase is simple to prepare, the analysis time is short, the analysis cost is greatly reduced, each impurity can be well separated and accurately located and calculated, and the quality control of fluticasone propionate aerosol can be provided, thereby ensuring the clinical efficacy.

[0053] Example 2 A UPLC detection kit for related substances of fluticasone propionate inhalation aerosol, comprising: a mobile phase A, which is 0.05% phosphoric acid water, obtained by adding 0.5 ml of 85% phosphoric acid to water to 1 L; a mobile phase B, which is acetonitrile; a diluent, which is prepared by mixing the mobile phase A and the mobile phase B according to a volume ratio of 1:1; impurity reference substances, including: impurity CCI11400 reference substance, impurity GR36264X reference substance, impurity GR269949X reference substance, impurity CCI22341 reference substance, impurity CCI18771 reference substance, impurity fluticasone propionate reference substance, impurity GR51610X reference substance, impurity GR40775X reference substance, impurity CCI18773 reference substance, impurity GR112801X reference substance, impurity high-temperature degradation reference substance, impurity GR68620X reference substance, impurity GR247095X reference substance; a specification for recording the UPLC conditions of the UPLC detection kit of the present embodiment, the UPLC conditions of which include: a detection wavelength of 239 nm, a flow rate of 0.60 ml / min, a column temperature of 45℃, an injection volume of 4 μL, and a gradient elution program as follows:

[0054] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. For those skilled in the art, the present application can have various modifications and changes. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. A UPLC analysis method for related substances in fluticasone propionate inhalation aerosol, characterized in that: The steps include: Acetonitrile and 0.030-0.070% by volume phosphoric acid water were mixed in a volume ratio of 1:0.9-1.1 to prepare a diluent, and the fluticasone propionate inhalation aerosol was dissolved in the diluent to prepare a sample solution. A chromatographic column filled with octadecylsilane bonded silica gel was used, with 0.030-0.070% by volume phosphoric acid water as mobile phase A and acetonitrile as mobile phase B. The sample solution was subjected to UPLC detection according to the gradient elution program shown in the table below; 。 2. The UPLC analysis method according to claim 1, wherein: The substances tested and analyzed include fluticasone propionate and fluticasone propionate aerosol impurities; the fluticasone propionate aerosol impurities are: 、 、 、 、 、 、 、 、 、 、 、 .

3. The UPLC analysis method according to claim 1, wherein: The gradient elution program is shown in the following table: 。 4. The UPLC analysis method according to claim 1, wherein the chromatographic column used in the UPLC detection is an ACQUITY UPLC BEH C18, 2.1 mm × 100 mm, 1.7 μm, or an equivalent performance chromatographic column.

5. The UPLC analysis method according to claim 1, wherein In UPLC detection, the detection wavelength is 229~249nm.

6. The UPLC analysis method according to claim 1, wherein In UPLC detection, the flow rate was 0.50~0.70ml / min.

7. The UPLC analysis method according to claim 1, wherein In UPLC detection, the column temperature is 40~50℃.

8. A UPLC detection kit for related substances of fluticasone propionate inhalation aerosol, characterized in that: include: The diluent is prepared by mixing acetonitrile and 0.030-0.070% by volume phosphoric acid water in a volume ratio of 1:0.9-1.1; Mobile phase: 0.030-0.070% volume percent phosphoric acid water as mobile phase A, acetonitrile as mobile phase B; A carrier for recording UPLC detection conditions; the UPLC detection conditions include a chromatographic column filler of octadecylsilane bonded silica gel and a gradient elution program; the gradient elution program is shown in the following table: 。 9. The UPLC detection kit according to claim 8, wherein Impurity controls are also included.

10. Use of the UPLC analysis method according to any one of claims 1 to 7 or the UPLC detection kit according to claim 8 or 9 for quality monitoring of related substances in fluticasone propionate inhalation aerosol during the production process or during the storage process of fluticasone propionate inhalation aerosol.

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