Determination method for content and dissolution rate of desmopressin acetate tablets

Abstract

The invention relates to the technical field of medicine analysis, in particular to a method for measuring the content and the dissolution rate of desmopressin acetate tablets, and the content and the dissolution rate of desmopressin acetate in the desmopressin acetate tablets are measured by adopting a high performance liquid chromatography. A ghost peak trapping small column is mounted in front of the sample injector; a chromatographic column filling agent is octadecyl silane bonded silica gel; the mobile phase A is a 0.1-0.3% phosphoric acid solution; the mobile phase B is acetonitrile; the diluent is an ammonium acetate solution; carrying out gradient elution; the flow rate is 0.9 to 1.1 ml per minute; the column temperature is 28-32 DEG C; and the sample injection volume is 80-100 [mu] l.

Description

Technical Field

[0001] The present application relates to the technical field of drug analysis, and in particular to a method for determining the content and dissolution of desmopressin acetate tablets. Background Art

[0002] Desmopressin Acetate Tablets can be used to treat central diabetes insipidus and nocturnal enuresis in patients aged six or above. The original manufacturer is Ferring (Switzerland) Pharmaceutical Co., Ltd. and it is now on the market.

[0003] Desmopressin Acetate Tablets are included in both the Chinese Pharmacopoeia and the British Pharmacopoeia. The consistency evaluation of generic drugs uses the reference preparation as a control to conduct formulation research, including comparative studies on prescription, impurities, content and dissolution curve of solid preparations, in order to improve the success rate of in vivo bioequivalence tests. Therefore, accurate and reliable detection methods are the prerequisite for the formulation of quality standards and provide a basis for the formulation of quality standards.

[0004] The structural formula of desmopressin acetate is shown below:

[0005] Summary of the invention

[0006] The purpose of the present invention is to establish a method for determining the content and dissolution of desmopressin acetate tablets. The present invention solves the technical problems of poor durability and low sensitivity caused by high salt concentration in the desmopressin acetate content detection method included in the Chinese Pharmacopoeia. The present determination method has high sensitivity and can accurately detect the content and dissolution of smaller-sized desmopressin acetate tablets (specification: 0.1 mg).

[0007] The present invention provides a method for determining the content and dissolution of desmopressin acetate tablets, comprising:

[0008] The content and solubility of desmopressin acetate in desmopressin acetate tablets were determined by HPLC.

[0009] A Welch Ghost-Buster Column 4.6mm×50mm was installed in front of the injector;

[0010] The column filler is octadecylsilane bonded silica gel;

[0011] Mobile phase A is 0.1-0.3% phosphoric acid solution; mobile phase B is acetonitrile; diluent is ammonium acetate solution; gradient elution is performed;

[0012] The flow rate is 0.9-1.1 ml per minute; the column temperature is 28-32°C; the injection volume is 80-100 μl.

[0013] In some embodiments, the detection wavelength for determining the desmopressin acetate content in the desmopressin acetate tablet is 235 nm.

[0014] In some embodiments, the detection wavelength for determining the solubility of desmopressin acetate in the desmopressin acetate tablet is 200 nm.

[0015] In some embodiments, the diluent ammonium acetate solution has a concentration of 0.025 mol / L.

[0016] In some embodiments, the strength of the desmopressin acetate tablet is 0.1 mg.

[0017] In some embodiments, the mobile phase A is a 0.2% phosphoric acid solution.

[0018] In some embodiments, the chromatographic column is Phenomenex Luna C18, 4.6 mm×30 mm, 5 μm.

[0019] In some embodiments, the flow rate is 1.0 ml per minute and the column temperature is 30°C.

[0020] In some embodiments, the injection volume is 100 μl.

[0021] In some embodiments, the gradient elution procedure is:

[0022] Table 1 Gradient elution program

[0023] Time (minutes) Mobile phase A (%) Mobile phase B (%) 0 90 10 4 62 38 4.1 90 10 7 90 10

[0024] Compared with the prior art, the beneficial effects of the present invention include:

[0025] The method for determining the content and dissolution of desmopressin acetate tablets provided by the present invention can accurately detect desmopressin acetate tablets of smaller specifications, simplifies the preparation of mobile phase A and diluent, reduces the analysis cost, and the recovery rate is between 99.9% and 102.1%; the system has good applicability, durability, linear relationship, sensitivity and accuracy. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to more clearly illustrate the technical solution of the present application, the drawings required for use in the embodiments are briefly introduced below. Obviously, for ordinary technicians in this field, other drawings can be obtained based on these drawings without any creative work.

[0027] Figure 1 This is the content determination spectrum of desmopressin acetate tablets;

[0028] Figure 2 This is the dissolution spectrum of desmopressin acetate tablets;

[0029] Figure 3 This is the content spectrum of desmopressin acetate tablets determined by the method recorded in the Chinese Pharmacopoeia;

[0030] Figure 4 This is the experimental spectrum for determining the durability of organic phase according to the method recorded in the Chinese Pharmacopoeia. DETAILED DESCRIPTION

[0031] In order to enable those skilled in the art to understand the characteristics and effects of the present invention, the following is a general description and definition of the terms and expressions mentioned in the specification and claims. Unless otherwise specified, all technical and scientific terms used in the text are the common meanings understood by those skilled in the art for the present invention. In the event of a conflict, the definition in this specification shall prevail.

[0032] The theories or mechanisms described and disclosed herein, whether correct or incorrect, should not limit the scope of the present invention in any way, that is, the present invention can be implemented without being limited by any specific theory or mechanism.

[0033] Herein, all features such as values, quantities, contents and concentrations defined in the form of numerical ranges or percentage ranges are for simplicity and convenience only. Accordingly, the description of numerical ranges or percentage ranges should be considered to have included and specifically disclosed all possible secondary ranges and individual values ​​within the range (including integers and fractions).

[0034] In this document, in order to make the description concise, not all possible combinations of various technical features in various embodiments or examples are described. Therefore, as long as there is no contradiction in the combination of these technical features, the various technical features in various embodiments or examples can be combined arbitrarily, and all possible combinations should be considered to be within the scope of this specification.

[0035] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention. In addition, it should be understood that after reading the content taught by the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms fall within the scope limited by the appended claims of the application equally.

[0036] The following examples use conventional instruments and equipment in the art. The experimental methods in the following examples where specific conditions are not specified are usually carried out under conventional conditions or under conditions recommended by the manufacturer. The various raw materials used in the following examples are conventional commercial products unless otherwise specified, and their specifications are conventional specifications in the art. In the specification of the present invention and the following examples, unless otherwise specified, "%" means weight percentage, "part" means weight part, and ratio means weight.

[0037] The following is a further description of the method for determining the content and dissolution of desmopressin acetate tablets provided by the present application in conjunction with specific examples.

[0038] Embodiment 1:

[0039] Desmopressin acetate tablets content determination method

[0040] 1. Chromatographic conditions

[0041] A Welch Ghost-Buster Column 4.6mm×50mm was installed in front of the injector;

[0042] Column: Phenomenex Luna C18, 4.6 mm × 30 mm, 5 μm;

[0043] Mobile phase: Mobile phase A is 0.2% phosphoric acid solution, mobile phase B is acetonitrile;

[0044] Preparation method of mobile phase: Mobile phase A is to measure 1000ml of water, accurately pipette 2ml of phosphoric acid, mix well, and degas by ultrasonication;

[0045] Mobile phase B is 1000 ml of acetonitrile, which is ultrasonically degassed;

[0046] Diluent: 0.025 mol / L ammonium acetate solution;

[0047] Preparation method of diluent: Take 1.93g of ammonium acetate, add 1000ml of water to dissolve, and degas by ultrasonication;

[0048] Detection wavelength: 235nm; flow rate: 1.0ml per minute; column temperature: 30℃;

[0049] Injection volume 100 μl; analysis time 7 min;

[0050] Elution method: gradient elution, elution program:

[0051] Table 1 Gradient elution program

[0052] Time (minutes) Mobile phase A (%) Mobile phase B (%) 0 90 10 4 62 38 4.1 90 10 7 90 10

[0053] 2. Solution preparation

[0054] Desmopressin acetate reference solution: Take about 20 mg of desmopressin acetate reference, accurately weigh it, place it in a 100 ml volumetric flask, add an appropriate amount of diluent and shake to dissolve, add diluent to the scale, shake well; accurately measure 5 ml of the solution, place it in a 100 ml volumetric flask, dilute it to the scale with diluent, and make up to volume. Prepare 2 portions in parallel (start from weighing).

[0055] Desmopressin acetate test solution: Take 10 tablets of this product, place in a 100ml volumetric bottle, add an appropriate amount of diluent (about 40-50% of the volume of the volumetric bottle), oscillate at 180-190 rpm for 10 minutes, add diluent to the scale, shake well, filter through a 0.45μm polyethersulfone filter membrane (or equivalent filter membrane), discard 1ml of the initial filtrate, and take the subsequent filtrate. Prepare 2 portions in parallel (starting from the preparation).

[0056] 3. Injection sequence

[0057] Table 2 Desmopressin acetate tablets content determination method injection sequence

[0058] 1 Blank solution (solvent) ≥1(until no residue*) 2 Reference solution 1 6 3 Reference solution 2 1 4 Test solution 1 2 5 Test solution 2 2 6 Reference solution 1 1

[0059] 4. The recovery rate of reference solution 2 and reference solution 1 is calculated as follows:

[0060]

[0061]

[0062]

[0063]

[0064] Where:

[0065] A RS —The average of the main peak areas of the reference solution 1 injected 6 times continuously;

[0066] A RS2 -The main peak area of ​​the reference solution 1;

[0067] C RS —Concentration of reference solution 1, mg / ml;

[0068] A CS -Main peak area of ​​reference solution 2;

[0069] C CS —Concentration of reference solution 2, mg / ml;

[0070] F—response factor of reference solution 1;

[0071] W S —Weighing amount of reference substance, mg;

[0072] H—reference substance content (content after deducting water and acetic acid), %;

[0073] A S —Average peak area of ​​main peak of reference solution 1;

[0074] D S—Dilution multiple of reference solution;

[0075] A t —The main peak area in the test solution;

[0076] D t —Dilution multiple of the test solution;

[0077] W t —Specifications of desmopressin tablets, 0.089 mg.

[0078] 5. Results

[0079] The content determination spectrum of the obtained desmopressin acetate tablets is as follows Figure 1 As shown, the peak time of desmopressin acetate is 3.75 min, the signal-to-noise ratio is 1114.8, and the peak width is 0.33. The method of the present invention can effectively avoid the peak interference of the auxiliary material povidone in the preparation, and has high sensitivity.

[0080] Embodiment 2:

[0081] Desmopressin acetate tablets dissolution test method

[0082] 1. Chromatographic conditions

[0083] A Welch Ghost-Buster Column 4.6mm×50mm was installed in front of the injector;

[0084] Column: Phenomenex Luna C18, 4.6 mm × 30 mm, 5 μm;

[0085] Mobile phase: Mobile phase A is 0.2% phosphoric acid solution, mobile phase B is acetonitrile;

[0086] Preparation method of mobile phase: Mobile phase A is to measure 1000ml of water, accurately pipette 2ml of phosphoric acid, mix well, and degas by ultrasonication;

[0087] Mobile phase B is 1000 ml of acetonitrile, which is ultrasonically degassed;

[0088] Diluent: 0.025 mol / L ammonium acetate solution;

[0089] Preparation method of diluent: Take 1.93g of ammonium acetate, add 1000ml of water to dissolve, and degas by ultrasonication;

[0090] Detection wavelength: 200nm; flow rate: 1.0ml per minute; column temperature: 30℃;

[0091] Injection volume 100 μl; analysis time 7 min;

[0092] Elution method: gradient elution, elution program:

[0093] Table 1 Gradient elution program

[0094] Time (minutes) Mobile phase A (%) Mobile phase B (%) 0 90 10 4 62 38 4.1 90 10 7 90 10

[0095] 2. Solution preparation

[0096] Diluent: Take 1.93 g of ammonium acetate, add 1000 ml of water to dissolve, filter, and degas by ultrasonication.

[0097] Medium: pH 4.5 acetate buffer.

[0098] pH 4.5 acetate buffer: weigh 2.99 g of sodium acetate trihydrate, dissolve it in water, add 14.0 ml of 2 mol / L acetic acid solution and dilute it to 1000 ml with water, adjust the pH to 4.5 with acetic acid.

[0099] Reference solution: Take about 20 mg of desmopressin acetate reference, weigh accurately, place in a 100 ml volumetric flask, add appropriate amount of diluent to dissolve, add diluent (0.025 mol / L ammonium acetate solution) to dilute to scale, shake well; accurately measure 1 ml of the solution, place in a 100 ml volumetric flask, dilute to scale with solvent, shake well; accurately measure 5 ml of the solution, place in a 50 ml volumetric flask, dilute to scale with medium (pH 4.5 acetate buffer), shake well, and you have it. Prepare 2 portions in parallel (start with weighing).

[0100] Test solution: Add 500 ml of dissolution medium to each of the 6 dissolution cups, take 6 tablets of this product, put them into each of the 6 dissolution cups, and perform the dissolution according to the above dissolution parameters. Take out 10 ml of the dissolution solution after 15 minutes, filter it through a 0.22 μm polyethersulfone filter membrane, discard 5 ml of the initial filtrate, and take the subsequent filtrate.

[0101] Note: The sample volume and volumetric bottle can be scaled up or down in the same proportion according to actual conditions.

[0102] 3. Injection sequence

[0103] Table 3 Desmopressin acetate tablets dissolution test method injection sequence

[0104]

[0105]

[0106] 4. The recovery rate of reference solution 2 and reference solution 1 is calculated as follows:

[0107]

[0108] The recovery rate of the returned reference solution 1 is calculated as follows:

[0109]

[0110] Dissolution curve calculation formula:

[0111]

[0112]

[0113]

[0114] Where:

[0115] A RS —The average of the main peak areas of the reference solution 1 injected 6 times in a row;

[0116] A RS2 -The main peak area of ​​the reference solution 1 is returned;

[0117] C RS —Concentration of reference solution 1, mg / ml;

[0118] A CS - Peak area of ​​main peak of reference solution 2;

[0119] C CS —Concentration of reference solution 2, mg / ml;

[0120] F—response factor of reference solution 1;

[0121] M RS —Weighing amount of reference substance, mg;

[0122] H—the content of reference substance, %;

[0123] Ds—dilution factor of reference solution, ml;

[0124] A 1 —The main peak area of ​​the test solution at -5 minutes;

[0125] A 2 —The main peak area of ​​the test solution at -30 minutes;

[0126] A 3 —The main peak area of ​​the test solution at -45 minutes;

[0127] W—labeled amount of test sample, mg.

[0128] 5. Results

[0129] Table 4 Desmopressin acetate tablets dissolution test results

[0130] Retention time (min) Peak area Theoretical plates Tailing Factor Peak width Signal-to-Noise Ratio (S / N) 3.745 77.45 9973 1.26 0.31 723.2

[0131] The dissolution spectrum of desmopressin acetate tablets obtained by determination is as follows Figure 2 As shown, the detection sensitivity can be improved by the method of the present invention.

[0132] Embodiment 3:

[0133] Investigation on the content determination method of desmopressin acetate tablets

[0134] (1) System suitability test

[0135] Table 5 Desmopressin acetate tablets content determination method system suitability experiment

[0136]

[0137] (2) Content durability test

[0138] Table 6 Desmopressin acetate tablets content determination method durability experiment

[0139]

[0140] (3) Content linearity experiment

[0141] Table 7 Desmopressin acetate tablets content determination method linear experiment

[0142]

[0143] (4) Content recovery experiment

[0144] Table 8 Desmopressin acetate tablets content determination method recovery experiment

[0145]

[0146] Embodiment 4:

[0147] Study on the dissolution determination method of desmopressin acetate tablets

[0148] (1) Dissolution linearity results

[0149] Table 9 Desmopressin acetate tablets dissolution determination method linear experiment

[0150]

[0151]

[0152] The linear range is 0.04μg / ml~0.33μg / ml.

[0153] (2) Dissolution accuracy results

[0154] Table 10 Desmopressin acetate tablets dissolution determination method accuracy experiment

[0155]

[0156] Comparative Example 1:

[0157] Chinese Pharmacopoeia Record Method-Content Determination Experiment

[0158] Table 11 Content determination experiment of the method recorded in Chinese Pharmacopoeia

[0159]

[0160] Comparative Example 2:

[0161] Chinese Pharmacopoeia Recording Method-Organic Phase Durability Test

[0162] like Figure 4 As shown, the proportion of the organic phase changed, the peak time and peak shape of desmopressin changed significantly, and the method was less robust.

[0163] Comparative Example 3:

[0164] Chinese Pharmacopoeia Record Method-Dissolution Linearity Test

[0165] Table 12 Dissolution linearity test of the method recorded in Chinese Pharmacopoeia

[0166]

[0167] The linear range is 0.1μg / ml~0.3μg / ml.

[0168] It can be seen from the results of the above embodiments and comparative examples that the method for determining the content and dissolution of desmopressin acetate tablets provided by the present invention can accurately detect desmopressin acetate tablets of smaller specifications, and the system has good applicability, durability, linear relationship, sensitivity and accuracy; the recovery rate is between 99.9% and 102.1%, the operation is simple, and the analysis time is short.

[0169] Although the present invention has been disclosed as above in the form of a preferred embodiment, it is not intended to limit the present invention. Anyone familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the definition of the claims.

Claims

1. A method for determining the content and dissolution of desmopressin acetate tablets, It is characterized in that include: The content and solubility of desmopressin acetate in desmopressin acetate tablets were determined by HPLC. Install a ghost peak capture column in front of the injector; The column filler is octadecylsilane bonded silica gel; Mobile phase A is 0.1-0.3% phosphoric acid solution; mobile phase B is acetonitrile; diluent is ammonium acetate solution; gradient elution is performed; The flow rate is 0.9-1.1 ml per minute; the column temperature is 28-32°C; the injection volume is 80-100 μl.

2. A method for determining the content and dissolution of desmopressin acetate tablets according to claim 1, Features: The detection wavelength for determining the desmopressin acetate content in desmopressin acetate tablets is 235 nm.

3. A method for determining the content and dissolution of desmopressin acetate tablets according to claim 1, Features: The detection wavelength for determining the solubility of desmopressin acetate in desmopressin acetate tablets is 200 nm.

4. A method for determining the content and dissolution of desmopressin acetate tablets according to claim 1, Features: The concentration of the diluent ammonium acetate solution is 0.025 mol / L.

5. A method for determining the content and dissolution of desmopressin acetate tablets according to claim 1, Features: The specification of desmopressin acetate tablets was determined to be 0.1 mg.

6. A method for determining the content and dissolution of desmopressin acetate tablets according to claim 1, Features: The mobile phase A is 0.2% phosphoric acid solution.

7. A method for determining the content and dissolution of desmopressin acetate tablets according to claim 1, Features: The ghost peak capture column is a Welch Ghost-Buster Column 4.6mm×50mm; the chromatographic column is a Phenomenex Luna C18, 4.6mm×30mm, 5μm.

8. A method for determining the content and dissolution of desmopressin acetate tablets according to claim 1, Features: The flow rate was 1.0 ml per minute and the column temperature was 30°C.

9. A method for determining the content and dissolution of desmopressin acetate tablets according to claim 1, Features: Injection volume: 100 μl.

10. A method for determining the content and dissolution of desmopressin acetate tablets according to claim 1, It is characterized in that The procedure of the gradient elution is: 。

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