Method for detecting content of povidone K30

Abstract

The invention belongs to the technical field of pharmaceutical analysis, and relates to a method for detecting the content of povidone K30. According to the method, the content of povidone K30 in the medicinal preparation is detected by adopting a high performance liquid chromatography; detection conditions of the high performance liquid chromatography are as follows: a chromatographic column is a C18-AQ column; a mobile phase is a phosphate solution. The method has the characteristics of high accuracy, strong specificity and the like for detecting the content of povidone K30 in the pharmaceutical preparation, can overcome the interference of other auxiliary materials in the pharmaceutical preparation, and has important application value in development and consistency evaluation of the pharmaceutical preparation.

Description

Technical Field

[0001] The invention belongs to the technical field of drug analysis, and particularly relates to a method for detecting the content of povidone K30. Background Art

[0002] Povidone K30 (PVP K30) is an organic compound with the molecular formula (C6H9NO) n , white to milky white powder; odorless or slightly odorous, tasteless. Its structural formula is as follows:

[0003]

[0004] Povidone K30 has a wide range of applications in medicine, among which the most widely used is as an adhesive for tablets and granules. PVP can also be used as a glidant for capsules, a detoxifying agent and lubricant for eye drops, a cosolvent for injections, a dispersant for liquid preparations, and a stabilizer for enzymes and thermosensitive drugs. Povidone can also be synthesized with iodine to form PVP-I disinfectant. In addition, PVP can also be used as a cryopreservative in medicine. There are hundreds of drugs that use PVP products as excipients. The development and consistency evaluation of generic drugs and the exemption of bioequivalence experiments all require consistency with the quality and efficacy of the original research. In order to achieve consistency with the quality and efficacy of the original research, reverse engineering technology came into being. As a commonly used excipient in pharmaceutical preparations, how to quickly detect its content in the preparation prescription is particularly important for generic drug development and consistency evaluation.

[0005] Chinese invention patent application CN107505424A discloses a HPLC method for determining the content of povidone in a povidone-iodine aqueous solution. In this method, a C18 chromatographic column is used, with an acetonitrile-water volume ratio of 5:95 as the mobile phase, a flow rate of 0.8mL / min, and an ultraviolet detection wavelength of 205nm to determine the content of povidone in a povidone-iodine aqueous solution. The test sample is analyzed and determined. This method cannot specifically determine the content of povidone K30, but can only determine the total amount of povidone. And the document "HPLC method for determining the content of povidone in povidone-iodine oral solution" also uses a C18 chromatographic column with an acetonitrile-water volume ratio of 5:95 as the mobile phase. However, the above detection methods are not suitable for the determination of the content of povidone K30 in oral solid preparations containing succinic acid excipients, and the excipients interfere with the detection of povidone K30.

[0006] Chinese invention patent application CN112684031A discloses a HPLC method for determining the content of povidone K30, in which a potassium nitrate-acetonitrile-water mixed solution is used as the mobile phase, and a molecular exclusion chromatography method is used to determine the content of povidone K30 in the product to be tested. Although the content of povidone K30 can be accurately determined, the chromatographic column is expensive, brittle, and the column is not tightly packed, resulting in low column efficiency.

[0007] Therefore, it is necessary to provide a HPLC determination method for the content of povidone K30 to solve the above-mentioned technical defects. Summary of the invention

[0008] In view of this, the object of the present invention is to provide a method for detecting the content of povidone K30 in an oral solid preparation containing succinic acid as an excipient. The method uses a conventional chromatographic column and is simple to operate. It provides supporting data for generic drug development, consistency evaluation, bioequivalence and other studies, and the content determination results are highly accurate and specific.

[0009] In order to achieve the above object, the present invention adopts the following technical solutions:

[0010] A method for detecting the content of povidone K30, which uses high performance liquid chromatography to detect the content of povidone K30 in a pharmaceutical preparation;

[0011] The detection conditions of the high performance liquid chromatography are as follows:

[0012] The chromatographic column is a C18-AQ column;

[0013] The mobile phase was phosphate solution.

[0014] Preferably, the C18-AQ column model is a Venusil MP C18 (2) column with specifications of inner diameter 4.6 mm×length 250 mm and particle size 5 μm.

[0015] Preferably, the phosphate in the phosphate solution is selected from one or more of potassium dihydrogen phosphate, dipotassium hydrogen phosphate, sodium dihydrogen phosphate and disodium hydrogen phosphate.

[0016] More preferably, the concentration of the phosphate solution is 0.01-0.1M, further preferably 0.05M.

[0017] More preferably, the pH of the phosphate solution is 2.0-4.0, and further preferably 2.38-2.42.

[0018] Preferably, the phosphate solution comprises a pH adjuster, and the pH adjuster is phosphoric acid.

[0019] Preferably, the detection conditions of the high performance liquid chromatography also include: elution mode: isocratic elution.

[0020] Preferably, the detection conditions of the high performance liquid chromatography also include: a detection wavelength of 195-215 nm, preferably 210 nm.

[0021] Preferably, the detection conditions of the high performance liquid chromatography method further include: a column temperature of 20-35°C, preferably 25°C.

[0022] Preferably, the detection conditions of the HPLC also include: a flow rate of 0.3-1.0 mL / min, preferably 0.5 mL / min.

[0023] Preferably, the detection conditions of the high performance liquid chromatography method also include: the injection volume is 10-30 μL, preferably 20 μL.

[0024] Preferably, the pharmaceutical preparation is a pharmaceutical preparation containing succinic acid as an excipient.

[0025] Preferably, the detection method specifically comprises the following steps:

[0026] (1) Prepare test sample solution and reference sample solution;

[0027] (2) Inject the test solution and the reference solution into a liquid chromatograph for testing;

[0028] (3) Calculate the content of povidone K30 in the drug preparation.

[0029] Further preferably, the preparation method of the test solution is: take the drug preparation, grind it into powder and weigh it accurately, add water and sonicate it, dilute it with water to make up the volume, centrifuge it, and take the supernatant, which is the test solution.

[0030] Further preferably, the preparation method of the reference substance solution is: accurately weigh povidone K30, place it in a volumetric bottle, add water and sonicate, dilute it with water to a fixed volume, and shake well to obtain the reference substance solution.

[0031] Further preferably, the present invention calculates the content of povidone K30 in the pharmaceutical preparation by an external standard method based on the chromatogram of the test solution and the chromatogram of the reference solution.

[0032] Specifically, the formula is used: result = (ru / rs) × (Cs / Cu) × 100, ru: peak area of ​​povidone K30 in the test solution, rs: peak area of ​​povidone K30 in the reference solution; Cs: concentration of povidone K30 in the reference solution (mg / mL); Cu: concentration of povidone K30 in the test solution (mg / mL).

[0033] By adopting the above chromatographic conditions, the content of povidone K30 in the pharmaceutical preparation can be accurately determined based on the peak area of ​​povidone K30 in the chromatogram with good accuracy. At the same time, the inventors also found that when other HPLC conditions are adopted, the characteristic peaks of excipients and povidone K30 may not be effectively separated, and thus the povidone K30 content of the pharmaceutical preparation cannot be accurately determined based on the peak area of ​​povidone K30 in the chromatogram.

[0034] Compared with the prior art, the present invention has the following beneficial effects:

[0035] The present invention has high accuracy and high specificity in detecting the content of povidone K30 in a pharmaceutical preparation. Neither the blank solution nor the blank excipient solution interferes with the determination of the content of povidone K30, and the content of each component in the pharmaceutical preparation can be accurately determined based on the peak area of ​​povidone K30 in the chromatogram. The method provides good technical support for the detection of povidone K30 in pharmaceutical preparations (especially pharmaceutical preparations containing succinic acid excipients), and has important application value in the development and consistency evaluation of pharmaceutical preparations. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 is the HPLC chromatogram of the test solution in Example 1, Figure 1 A in the figure is the complete map, and B is the local enlarged map;

[0037] Figure 2 is the HPLC chromatogram of the blank solution in Experimental Example 2, Figure 2 A in the figure is the complete map, and B is the local enlarged map;

[0038] Figure 3 is the HPLC chromatogram of the blank auxiliary material solution in Experimental Example 2, Figure 3 A in the figure is the complete map, and B is the local enlarged map;

[0039] Figure 4 This is a high performance liquid chromatogram overlay comparison diagram of the blank excipient solution and the reference substance solution in Experimental Example 2. Figure 4 A in the figure is the complete map, and B is the local enlarged map;

[0040] Figure 5 This is a high performance liquid chromatogram overlay comparison of the reference solution and the test solution in Comparative Example 1. Figure 5 A in the figure is the complete map, and B is the local enlarged map;

[0041] Figure 6 This is a high performance liquid chromatogram overlay comparison of the reference solution and the test solution in Comparative Example 2. Figure 6 A in the figure is the complete map, and B is the local enlarged map;

[0042] Figure 7 This is a high performance liquid chromatogram overlay comparison of the reference solution and the test solution in Comparative Example 3. Figure 7 A in the figure is the complete map, and B is the local enlarged map;

[0043] Figure 8 This is a high performance liquid chromatogram overlay comparison of the reference solution and the test solution in Comparative Example 4. Figure 8 A is the complete map, and B is the local enlarged map. DETAILED DESCRIPTION

[0044] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0045] In order to further illustrate the present invention, the following examples are used to describe the present invention in detail. The raw materials and auxiliary materials used in the following examples of the present invention are all commercially available products.

[0046] Instruments: Shimadzu LC-20AT high performance liquid chromatograph; Shimadzu AUW120D 1 / 100,000 electronic balance;

[0047] Test drugs and reagents: ferrous succinate preparation, povidone K30 (source: Anhui Shanhe Yaoyong Auxiliary Materials Co., Ltd.); potassium dihydrogen phosphate (Guangzhou Chemical Reagent Factory), phosphoric acid (Guangzhou Chemical Reagent Factory), succinic acid (source: Jiudian Hongyang Pharmaceutical Co., Ltd.), calcium hydrogen phosphate dihydrate (source: Hunan Jiudian Pharmaceutical Co., Ltd.), corn starch (source: Anhui Shanhe Yaoyong Auxiliary Materials Co., Ltd.), magnesium stearate (source: Anhui Shanhe Yaoyong Auxiliary Materials Co., Ltd.), ferrous succinate (source: Hunan Jiudian Pharmaceutical Co., Ltd.).

[0048] Example 1

[0049] The determination method of the content of povidone K30 in the pharmaceutical preparation (ferrous succinate preparation) of the present embodiment is as follows:

[0050] (1) Test solution: Grind 10 tablets of ferrous succinate into powder, accurately weigh 200 mg, place in a 20 mL volumetric flask, add water to 2 / 3 of the flask, sonicate for 15 min, cool to room temperature, dilute to scale with water, shake well, centrifuge, and take the supernatant as the test solution.

[0051] (2) Reference solution: Take 25 mg of povidone K30, weigh accurately, place in a 250 mL volumetric flask, add water to 2 / 3 of the flask, sonicate for 15 min, cool to room temperature, dilute to the mark with water, shake well, and use as the reference solution.

[0052] (3) The test solution and the reference solution are tested respectively by high performance liquid chromatography to obtain a chromatogram of the test solution and a chromatogram of the reference solution, wherein the chromatographic conditions are as follows:

[0053] The chromatographic column is a chromatographic column filled with octadecylsilane bonded silica gel (Venusil MpC18(2), 4.6mm*250mm5μm); the mobile phase is 0.05mol / L potassium dihydrogen phosphate solution (the pH is adjusted to 2.40 with phosphoric acid); the detection wavelength is 210nm; the injection volume is 20μL; the column temperature is 20℃; and the flow rate is 0.5mL / min.

[0054] (4) Based on the chromatograms of the test solution and the reference solution, Figure 1 As shown, Figure 1 A is the complete spectrum, and B is a partial enlarged view. The content of povidone K30 in the drug preparation was calculated using the external standard method, and the result was 1.03%.

[0055] Example 2

[0056] This example verifies the accuracy of the HPLC method of the present invention by specificity. The determination method is respectively determined according to the description of the experimental method in the above Example 1, and the determination results are shown in Table 1. The solution is prepared as follows:

[0057] (1) Blank solution: water.

[0058] (2) Blank excipient solution: Weigh 1.5 g of ferrous succinate, 1.5 g of succinic acid, 0.8 g of calcium hydrogen phosphate dihydrate, 1.4 g of corn starch, and 0.08 g of magnesium stearate, and mix them evenly. Take 200 mg of the mixture, weigh it accurately, place it in a 20 mL volumetric flask, add water to 2 / 3 of the volumetric flask, sonicate for 15 min, cool to room temperature, dilute with water to the scale, shake well, centrifuge, and take the supernatant as the blank excipient solution.

[0059] (3) Reference solution: Take 25 mg of povidone K30, weigh accurately, place in a 250 mL volumetric flask, add water to 2 / 3 of the flask, sonicate for 15 min, cool to room temperature, dilute with water to the mark, shake well, and use as the reference solution.

[0060] (4) The blank solution, blank auxiliary material solution and reference substance solution were respectively injected into a high performance liquid chromatograph and tested using the high performance liquid chromatography conditions of Example 1. The results are shown in FIG. Figure 2-Figure 4 .

[0061] Among them, the chromatogram in the blank solution is as follows Figure 2 As shown, Figure 2 A is the complete spectrum, and B is the partial enlarged view. The chromatogram of the blank auxiliary material solution is as follows Figure 3 As shown, Figure 3 A in the figure is the complete spectrum, and B is the partial enlarged view. In addition, the comparative chromatograms of the blank excipient solution and the reference substance solution are shown in Figure 4 As shown, Figure 4A in the figure is the complete spectrum, and B is the partial enlarged picture. It can be seen that the blank solution and the blank excipient solution do not interfere with the content determination of povidone K30.

[0062] Example 3

[0063] This example verifies the accuracy of the HPLC method of the present invention through a recovery experiment. Three levels of test sample solutions, 50%, 100%, and 150%, were selected and the recovery rates were measured respectively according to the HPLC conditions of Example 1. The results are shown in Table 1.

[0064] Table 1 Recovery results of polyvinylpyrrolidone K30

[0065]

[0066] It can be seen from the above table that the recovery rate RSD of the content determination of PVP K30 using the HPLC conditions of the present invention is less than 3%, and the content of each component in the pharmaceutical preparation can be accurately determined based on the peak area of ​​PVP K30 in the HPLC chromatogram.

[0067] Example 4

[0068] This example verifies the stability of the test solution of the oral solid preparation containing povidone K30. The content of povidone K30 in the test solution was measured 4 hours and 8 hours after preparation according to the HPLC conditions of Example 1. The results are shown in Table 2.

[0069] Table 2

[0070] content(%) Absolute value of the difference from 0h (%) 0h 1.09 / 4h 1.15 0.06 8h 1.13 0.04

[0071] It can be seen from the above table that the test solution of the oral solid preparation containing povidone K30 obtained by the method of the present invention has good stability.

[0072] Comparative Example 1

[0073] In this comparative example, reference is made to patent application number CN201610423183.7—HPLC determination method of povidone content in povidone iodine aqueous solution, and the determination method of document "HPLC determination of povidone content in povidone iodine oral solution", and the HPLC conditions are as follows: chromatographic column: Agilent Eclipse Plus C18 4.6mm×150mm, 5μm; detection wavelength: 205nm; column temperature: 30°C; flow rate 0.8ml / min; injection volume: 20μL; acetonitrile-water 5:95 as the mobile phase.

[0074] The specific operations are as follows:

[0075] (1) Test solution: Grind 10 tablets of ferrous succinate preparation into powder, take out 200 mg, accurately weigh, and place in a 20 mL volumetric flask. Add water to 2 / 3 of the volumetric flask, sonicate for 15 min, cool to room temperature, dilute to the mark with water, shake well, centrifuge, and take the supernatant as the test solution.

[0076] (2) Reference solution: Take 25 mg of povidone K30, weigh accurately, place in a 250 mL volumetric flask, add water to 2 / 3 of the flask, sonicate for 15 min, cool to room temperature, dilute to the mark with water, shake well, and use as the reference solution.

[0077] (3) The test solution and the reference solution were injected into the high performance liquid chromatograph for testing. Figure 5 As shown, Figure 5 A in the figure is the complete spectrum, and B is the local enlarged picture; it can be seen that the peak of povidone K30 in the test solution has a shoulder peak that is not completely separated, which interferes with the determination of the content of povidone K30 to a certain extent.

[0078] Comparative Example 2

[0079] In this comparative example, reference is made to patent application number CN201811055307.6—Determination of N-methylpyrrolidone and povidone K30 residues in hemodialyzers. The HPLC conditions are as follows: chromatographic column: Agilent Eclipse Plus C18 4.6 mm × 150 mm, 5 μm; detection wavelength: 205 nm; column temperature: 30 ° C; flow rate 0.8 mL / min; injection volume: 20 μL; acetonitrile-buffer solution (0.05 mol / L potassium dihydrogen phosphate, pH adjusted to 2.40 with phosphoric acid) 5:95 as the mobile phase.

[0080] The specific operations are as follows:

[0081] (1) Test solution: Grind 10 tablets of ferrous succinate preparation into powder, take out 200 mg, accurately weigh, and place in a 20 mL volumetric flask. Add water to 2 / 3 of the volumetric flask, sonicate for 15 min, cool to room temperature, dilute to the mark with water, shake well, centrifuge, and take the supernatant as the test solution.

[0082] (2) Reference solution: Take 25 mg of povidone K30, weigh accurately, place in a 250 mL volumetric flask, add water to 2 / 3 of the flask, sonicate for 15 min, cool to room temperature, dilute to the mark with water, shake well, and use as the reference solution.

[0083] (3) The test solution and the reference solution were injected into the high performance liquid chromatograph for testing. Figure 6 As shown, Figure 6A is the complete spectrum, and B is a local enlarged view; it can be seen that the main peak in the test solution is not completely separated from povidone K30, which obviously interferes with the determination of the content of povidone K30.

[0084] Comparative Example 3

[0085] The difference between this comparative example and Example 1 is that the mobile phase is different. This comparative example uses acetonitrile-buffer solution (0.05 mol / L potassium dihydrogen phosphate, pH adjusted to 2.40 with phosphoric acid) 5:95 as the mobile phase.

[0086] The specific operations are as follows:

[0087] (1) Test solution: Grind 10 tablets of ferrous succinate preparation into powder, take out 200 mg, accurately weigh, and place in a 20 mL volumetric flask. Add water to 2 / 3 of the volumetric flask, sonicate for 15 min, cool to room temperature, dilute to the mark with water, shake well, centrifuge, and take the supernatant as the test solution.

[0088] (2) Reference solution: Take 25 mg of povidone K30, weigh accurately, place in a 250 mL volumetric flask, add water to 2 / 3 of the flask, sonicate for 15 min, cool to room temperature, dilute to the mark with water, shake well, and use as the reference solution.

[0089] (3) The test solution and the reference solution were injected into the high performance liquid chromatograph for testing. Figure 7 As shown, Figure 7 A in the figure is a complete spectrum, and B is a local enlarged view; it can be seen that after the mobile phase of the present invention is replaced by the above mobile phase, there is a significant interference in the content determination of povidone K30.

[0090] Comparative Example 4

[0091] The difference between this comparative example and Example 1 is that the chromatographic column is different. The chromatographic column of this comparative example is a C18-AQ column (Agilent Eclipse Plus C18, 4.6 mm*250 mm 5 μm).

[0092] The specific operations are as follows:

[0093] (1) Test solution: Grind 10 tablets of ferrous succinate preparation into powder, take out 200 mg, accurately weigh, and place in a 20 mL volumetric flask. Add water to 2 / 3 of the volumetric flask, sonicate for 15 min, cool to room temperature, dilute to the mark with water, shake well, centrifuge, and take the supernatant as the test solution.

[0094] (2) Reference solution: Take 25 mg of povidone K30, weigh accurately, place in a 250 mL volumetric flask, add water to 2 / 3 of the flask, sonicate for 15 min, cool to room temperature, dilute to the mark with water, shake well, and use as the reference solution.

[0095] (3) The test solution and the reference solution were injected into the high performance liquid chromatograph for testing. Figure 8 As shown, Figure 8 A is the complete spectrum, and B is a partial enlarged view; it can be seen that the separation effect is also good when the chromatographic column of the present invention is replaced by Eclipse Plus C18, but it is slightly worse than Venusil MP C18 (2) of the present invention. When the C18-AQ chromatographic column is used to detect the content of povidone K30 in the pharmaceutical preparation, Venusil MP C18 (2) is preferred.

[0096] The above detailed description is a specific description of one feasible embodiment of the present invention. The embodiment is not intended to limit the patent scope of the present invention. Any equivalent implementation or modification that does not deviate from the present invention should be included in the scope of the technical solution of the present invention.

Claims

1. A method for detecting the content of povidone K30, characterized in that: The content of povidone K30 in the drug preparation was detected by high performance liquid chromatography; The detection conditions of the high performance liquid chromatography are as follows: The chromatographic column is a C18-AQ column; The mobile phase was phosphate solution.

2. The detection method according to claim 1, characterized in that: The C18-AQ column model is Venusil MP C18 (2), with specifications of inner diameter 4.6 mm×length 250 mm and particle size 5 μm.

3. The detection method according to claim 1, characterized in that: The phosphate in the phosphate solution is selected from one or more of potassium dihydrogen phosphate, dipotassium hydrogen phosphate, sodium dihydrogen phosphate and disodium hydrogen phosphate; The phosphate solution has a concentration of 0.01-0.1 M and a pH of 2.0-4.

0.

4. The detection method according to claim 1, characterized in that: The phosphate solution includes a pH adjuster, and the pH adjuster is phosphoric acid.

5. The detection method according to claim 1, characterized in that: The detection conditions of the high performance liquid chromatography also include: the elution method is isocratic elution.

6. The detection method according to claim 1, characterized in that: The detection conditions of the high performance liquid chromatography also include: the detection wavelength is 195-215nm.

7. The detection method according to claim 1, characterized in that: The detection conditions of the high performance liquid chromatography also include: the column temperature is 20-35°C.

8. The detection method according to claim 1, characterized in that: The detection conditions of the high performance liquid chromatography method also include: a flow rate of 0.3-1.0 mL / min and an injection volume of 10-30 μL.

9. The detection method according to claim 1, characterized in that: The pharmaceutical preparation is a pharmaceutical preparation containing succinic acid as an auxiliary material.

10. The detection method according to any one of claims 1 to 9, characterized in that: The detection method specifically comprises the following steps: (1) Prepare test sample solution and reference sample solution; (2) Inject the test solution and the reference solution into a high performance liquid chromatograph for testing; (3) Calculate the content of povidone K30 in the drug preparation based on the chromatogram peak area.

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