A method for detecting matrix components in miconazole nitrate suppositories

Abstract

The invention discloses a method for detecting matrix components in a miconazole nitrate suppository, which adopts HPLC-ELSD detection and specifically comprises the following steps: 1) preparing a reference solution; 2) preparing a test solution; 3) respectively drawing the reference solution and the test solution and injecting them into a high performance liquid chromatograph. The method for detecting matrix components in the miconazole nitrate suppository of the invention completely separates the matrix components of the miconazole nitrate suppository in a chromatogram through a specific sample pretreatment method and chromatographic conditions, and provides a new method for analyzing the composition of the matrix components of the miconazole nitrate suppository.

Description

Technical Field

[0001] The invention specifically relates to a method for detecting matrix components in miconazole nitrate suppositories. Background Art

[0002] Miconazole nitrate suppository is an antibacterial drug for external use in gynecology. It has a broad-spectrum antifungal effect, can affect the permeability of fungal cell membranes, and inhibit fungal growth. It has a strong antibacterial effect on Candida albicans and some Gram-positive bacteria, and can kill bacteria and relieve itching. It is an over-the-counter drug for local treatment of Candida vulvovaginal disease and severe infections caused by Gram-positive bacteria.

[0003] At present, the prescription of miconazole nitrate suppository preparation includes the main drug miconazole nitrate and a fatty matrix mixed fatty acid glyceride. The composition of the matrix mixed fatty acid glyceride directly affects the release of the drug. However, there is no analytical method for the composition of the matrix component in miconazole nitrate suppository, and the domestic and international implementation standards do not include relevant inspection items. As a result, the efficacy of the application varies greatly due to the uncontrollable matrix composition, and quality problems cannot be traced. Summary of the invention

[0004] In order to solve the above problems, the present invention provides a method for detecting matrix components in miconazole nitrate suppositories, which adopts HPLC-ELSD detection and specifically comprises the following steps:

[0005] 1) Preparation of reference solution: Take miconazole nitrate reference substance, add methanol-tetrahydrofuran to dissolve it, and use it as reference solution;

[0006] 2) Preparation of test solution: melt miconazole nitrate suppositories, condense them, and add methanol-tetrahydrofuran water bath until the oil layer is clear, which is used as the test solution;

[0007] 3) Pipette the reference solution and the test solution into the HPLC respectively; the chromatographic conditions are as follows:

[0008] Chromatographic column: octadecylsilane bonded silica gel as filler; mobile phase: acetonitrile-acetic acid as mobile phase A, dichloromethane as mobile phase B; gradient elution program: 0-4 min, 0% B; 4-12 min, 0% B→30% B; 12-18 min, 30% B; 18-30 min, 30% B→70% B; 30-35 min, 70% B; 35-36 min, 70% B→0% B; 36-45 min, 0% B.

[0009] Furthermore, the reference solution in step 1) contains 10 to 100 μg of miconazole nitrate per 1 ml.

[0010] Furthermore, in step 2), the mass volume ratio of the miconazole nitrate suppository to methanol-tetrahydrofuran is 0.1-0.5 g:100 ml, preferably 0.12:100 ml.

[0011] Furthermore, in step 2), the volume ratio of methanol to tetrahydrofuran is 1 to 3:1, preferably 1:1.

[0012] Furthermore, the chromatographic column is Gemini C18 110A, 4.6×250 mm, 5 μm, phenomenex.

[0013] Furthermore, the mobile phase A is acetonitrile-acetic acid in a volume ratio of 100:0.5.

[0014] Furthermore, the chromatographic conditions also include a column temperature of 30° C., a flow rate of 1 ml / min, and an injection volume of 5 μl.

[0015] Furthermore, the chromatographic conditions also include a drift tube temperature of 85° C. for the evaporative light scattering detector, a gas flow rate of 2.5 L / min, a Gain value of 1, and a non-split mode.

[0016] Furthermore, the chemical formula of the matrix component is C 35 H 66 O6, C 37 H 70 O6, C 39 H 74 O6, C 41 H 78 O6, C 43 H 82 O6, C 45 H 86 O6, C 47 H 90 O6, C 49 H 94 O6, C 51 H 98 O6, C 53 H 102 O6, C 55 H 106 O6 and / or C 57 H 110 O6.

[0017] Furthermore, the chromatographic peak characteristic parameters of the matrix component are as follows: Figure 1 shown.

[0018] The detection method of the matrix components in the miconazole nitrate suppository of the present invention completely separates the matrix components of the miconazole nitrate suppository in the chromatogram through a specific sample pretreatment method and chromatographic conditions, thus providing a new method for analyzing the composition of the matrix components of the miconazole nitrate suppository.

[0019] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0020] The above contents of the present invention are further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 Typical chromatogram (1: Miconazole nitrate C 18 H 14 Cl4N2O·HNO3,2:C 35 H 66 O6, 3: C 37 H 70 O6, 4: C 39 H 74 O6, 5: C 41 H 78 O6, 6: C 43 H 82 O6, 7: C 45 H 86 O6, 8: C 47 H 90 O6, 9: C 49 H 94 O6,

[0022] 10: C 51 H 98 O6, 11: C 53 H 102 O6, 12: C 55 H 106 O6, 13: C 57 H 110 O6) DETAILED DESCRIPTION

[0023] Example 1: Detection of matrix components in miconazole nitrate suppositories of the present invention

[0024] 1. Preparation of test solution

[0025] Take 5 miconazole nitrate suppositories, melt them at a low temperature, mix them evenly, and after condensation, accurately weigh 0.12 g, place it in a 100 ml volumetric flask, add methanol-tetrahydrofuran (1:1) mixed solvent, bathe in warm water and shake from time to time to clarify the oil layer, cool, dilute to the scale with the mixed solvent, shake well, and use it as the test solution.

[0026] 2. Preparation of reference solution

[0027] Take miconazole nitrate reference substance and dilute it with methanol-tetrahydrofuran (1:1) mixed solvent to make a reference substance solution containing about 50 μg per 1 ml.

[0028] 3. HPLC analysis conditions

[0029] The chromatographic column was Gemini C18 110A (4.6×250mm, 5μm, phenomenex), the mobile phase A was acetonitrile-acetic acid (100:0.5), the mobile phase B was dichloromethane, the column temperature was 30°C, the flow rate was 1ml / min; the evaporative light scattering detector (ELSD) was used: the drift tube temperature was 85°C, the gas flow rate was 2.5L / min, the Gain value was 1, and the non-split mode was used. Accurately measure 5μl of the test solution and inject it into the chromatograph to record the chromatogram.

[0030] The gradient elution program was:

[0031]

[0032] The beneficial effects of the present invention are described below through test examples.

[0033] Test Example 1 Method Precision

[0034] According to the detection method of Example 1, under the same experimental conditions, the same test sample was continuously measured three times, and the peak area RSD of each component was calculated. The results are shown in Table 1. As can be seen from Table 1, the relative standard deviation (RSD) of the mixed fatty acid glyceride compound (2-13) in the parallel determination results is 1.1% to 4.9%, indicating that the method has good repeatability and can meet the requirements for repeatability of matrix component detection in miconazole nitrate suppositories.

[0035] Table 1 Results of method precision investigation

[0036]

[0037]

[0038] In summary, the present invention completely separates the matrix components of miconazole nitrate suppositories in the chromatogram through a specific sample pretreatment method and chromatographic conditions, thereby providing a new method for analyzing the composition of the matrix components of miconazole nitrate suppositories.

Claims

1. A method for detecting matrix components in miconazole nitrate suppositories, characterized in that: It is detected by HPLC-ELSD, which includes the following steps: 1) Preparation of reference solution: Take miconazole nitrate reference substance and dissolve it in methanol-tetrahydrofuran to prepare the reference solution; 2) Preparation of test solution: melt miconazole nitrate suppositories, condense them, and add methanol-tetrahydrofuran water bath until the oil layer is clear, which is used as the test solution; 3) Pipette the reference solution and the test solution into the HPLC respectively; the chromatographic conditions are as follows: Chromatographic column: octadecylsilane bonded silica gel as filler; mobile phase: acetonitrile-acetic acid with a volume ratio of 100:0.5 as mobile phase A and dichloromethane as mobile phase B; gradient elution program: 0-4 min, 0% B; 4-12 min, 0% B→30% B; 12-18 min, 30% B; 18-30 min, 30% B→70% B; 30-35 min, 70% B; 35-36 min, 70% B→0% B; 36-45 min, 0% B; The chemical formula of the matrix component is C 35 H 66 O6, C 37 H 70 O6, C 39 H 74 O6, C 41 H 78 O6, C 43 H 82 O6, C 45 H 86 O6, C 47 H 90 O6, C 49 H 94 O6, C 51 H 98 O6, C 53 H 102 O6, C 55 H 106 O6 and C 57 H 110 O6.

2. The detection method according to claim 1, characterized in that: Step 1) The reference solution contains 10-100 μg of miconazole nitrate per 1 ml.

3. The detection method according to claim 1, characterized in that: Step 2) The mass volume ratio of the miconazole nitrate suppository to methanol-tetrahydrofuran is 0.1-0.5 g:100 ml.

4. The detection method according to claim 3, characterized in that: Step 2) The mass volume ratio of the miconazole nitrate suppository to methanol-tetrahydrofuran is 0.12:100 ml.

5. The detection method according to claim 1, 3 or 4, characterized in that: Step 2) The volume ratio of methanol to tetrahydrofuran is 1-3:

1.

6. The detection method according to claim 5, characterized in that: Step 2) The volume ratio of methanol to tetrahydrofuran is 1:

1.

7. The detection method according to claim 1, characterized in that: The chromatographic column is Gemini C18 110A, 4.6×250 mm, 5 μm, phenomenex.

8. The detection method according to claim 1, characterized in that: The chromatographic conditions also include a column temperature of 30° C., a flow rate of 1 ml / min, and an injection volume of 5 μl.

9. The method according to claim 6, characterized in that: The chromatographic conditions also include a drift tube temperature of 85° C. for the evaporative light scattering detector, a gas flow rate of 2.5 L / min, a gain value of 1, and a non-split mode.

Images