Method for determining and analyzing dissolution curve of tacrolimus capsule

Abstract

The invention discloses a method for determining and analyzing a dissolution curve of a tacrolimus capsule, which is characterized in that a high performance liquid chromatograph is adopted for determination, and a chromatographic column is a Prodigy Phenyl-3 phenyl column; a mobile phase is composed of acetonitrile, water, methanol and dilute phosphoric acid, and the ratio of the mobile phase is 460: 360: 180: 1; the pH of the dissolution medium is 6.8, and the pH of the dissolution medium is 4.5; the invention also provides preparation of a test solution and a reference solution. The high performance liquid chromatography analysis method of the tacrolimus capsule provided by the invention is suitable for large-scale detection, and the detection efficiency is greatly improved; the method also has the advantages of simplicity in operation, high accuracy, high precision and good stability.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical analysis technology, and more specifically to a method for determining the dissolution curve of tacrolimus capsules. Background Technology

[0002] Tacrolimus is a fermentation product isolated from Streptomyces. It is a macrolide drug and a potent immunosuppressant that is widely used in organ transplantation. However, it has the characteristics of "low water solubility and high permeability", so dissolution curve testing is required to ensure that the drug dissolves effectively in the body, thereby avoiding differences in efficacy due to dissolution problems.

[0003] In order to reduce the analytical costs of manufacturers, continuously improve the quality standards of this type of drug, thereby improving drug quality and further ensuring the safety of medication for the general public, there is an urgent need to provide a new analytical method for determining the dissolution curve of tacrolimus capsules. Summary of the Invention

[0004] This invention provides a method for determining the dissolution curve of tacrolimus capsules. This method is low in cost, has good separation effect, strong specificity, and is automated, thus ensuring the quality control of tacrolimus capsules.

[0005] Specifically, this invention discloses a method for determining the dissolution curve of tacrolimus capsules, which uses high performance liquid chromatography (HPLC) for determination.

[0006] The chromatographic column was a Prodigy Phenyl-3 phenyl column; the mobile phase consisted of acetonitrile-water-methanol-dilute phosphoric acid in a ratio of 460:360:180:1.

[0007] Dissolution medium: pH 6.8, pH 4.5.

[0008] Preparation of the test solution: Take tacrolimus capsules and sample them at 5 min, 10 min, 15 min, 20 min, 30 min, 45 min, 60 min, 90 min, and 120 min. Filter the samples through a 0.45 μm microporous membrane, take 5 ml of the filtrate, add 2 ml of acetonitrile and mix well to obtain the final product. Preparation of the reference solution: Accurately weigh an appropriate amount of tacrolimus reference standard, dissolve and dilute it in acetonitrile to prepare a solution containing approximately 0.015 mg per ml. Take 2 ml of the solution, add dissolution medium and mix well to obtain the final product. Take 400 μl of each of the test solution and the reference solution and inject them into the liquid chromatograph. Record the chromatograms and calculate the dissolution rate at each time point using the peak area according to the external standard method.

[0009] Preferably, the chromatographic conditions are: column temperature 45℃; flow rate 1.0 ml / min; detection wavelength 210 nm.

[0010] Preferably, the chromatograph is a Shimadzu high-performance liquid chromatograph. Preferably, the dissolution speed of the dissolution medium is 50 rpm or 75 rpm.

[0011] Compared with the prior art, the beneficial effects of the present invention are: The high-performance liquid chromatography method for the determination and analysis of tacrolimus capsules provided by this invention is suitable for large-scale detection and greatly improves detection efficiency.

[0012] Secondly, this method has the advantages of being simple to operate, highly accurate, highly precise, and stable. Attached Figure Description

[0013] Figure 1 The chromatogram of the blank solution in this invention is shown; Figure 2 The chromatogram of the tacrolimus reference solution is shown; Figure 3 The chromatogram of the tacrolimus capsule test solution is shown. Detailed Implementation

[0014] The present invention will now be described in detail with reference to various embodiments. However, it should be noted that these embodiments are not intended to limit the present invention. Equivalent changes or substitutions in function, method, or structure made by those skilled in the art based on these embodiments are all within the protection scope of the present invention.

[0015] In this invention, the term "blank solution" refers to a solution used in high-performance liquid chromatography (HPLC) detection and analysis methods, prepared under conditions identical to those used when measuring the sample, to eliminate interference. The measured result is called the "blank value," which should be subtracted from the sample measurement result to improve the accuracy of the determination.

[0016] In this invention, the term "test solution" refers to the solution of the sample to be tested in a high-performance liquid chromatography (HPLC) detection and analysis method.

[0017] In this invention, the term "reference solution" refers to a solution of a standard sample used in high-performance liquid chromatography (HPLC) detection and analysis methods.

[0018] In this invention, the term "mobile phase" refers to the substance that carries the analyte forward during liquid chromatography.

[0019] This invention discloses a method for determining and analyzing the dissolution curve of tacrolimus capsules, which is performed by high performance liquid chromatography.

[0020] The chromatographic conditions for the above detection method are as follows: Instruments used: Shimadzu high performance liquid chromatograph; Chromatographic column: Prodigy Phenyl-3 (pH-3) phenyl column (4.6*250mm, 5μm).

[0021] The mobile phase was acetonitrile-water-methanol-dilute phosphoric acid (3→50) (460:360:180:1).

[0022] The blank solution serves as the dissolution medium.

[0023] Dissolution media: pH 6.8, pH 4.5; dissolution speed of the dissolution media is 50 rpm, 75 rpm.

[0024] Column temperature 45℃; flow rate 1.0 ml / min; detection wavelength 210 nm.

[0025] The injection volume was 400 μl.

[0026] Experimental steps: Solution preparation: (1) Preparation of reference solution: Accurately weigh an appropriate amount of tacrolimus reference standard, dissolve and dilute it in acetonitrile to prepare a solution containing approximately 0.015 mg per ml. Take 2 ml of the solution and add 5 ml of dissolution medium, then mix well.

[0027] (2) Preparation of the test solution: Take tacrolimus capsules and sample them according to the first method of General Chapter 0931 of the Chinese Pharmacopoeia, at 5 min, 10 min, 15 min, 20 min, 30 min, 45 min, 60 min, 90 min, and 120 min. Filter the samples through a 0.45 μm microporous membrane, take 5 ml of the filtrate, add 2 ml of acetonitrile and mix well to obtain the final product. Accurately pipette 400 μl each of the test solution and the reference solution, inject them into the liquid chromatograph, record the chromatograms, and calculate the dissolution rate at each time point using the peak area according to the external standard method.

[0028] Figure 1 This is the chromatogram of the blank solution.

[0029] Figure 2 This is the chromatogram of the tacrolimus reference solution.

[0030] Figure 3 Chromatogram of the tacrolimus capsule test solution.

[0031] The results showed that the blank solvent did not interfere with the detection of the tacrolimus capsule dissolution curve. The chromatogram of the reference solution showed good peak shape and purity, with a tailing factor not exceeding 1.5, meeting the standard requirements.

[0032] In summary, the high-performance liquid chromatography method for the determination and analysis of tacrolimus capsules provided by this invention is suitable for large-scale detection and greatly improves detection efficiency.

[0033] The detection and analysis method provided by this invention also has the advantages of simple operation, high accuracy, high precision, and good stability.

[0034] The detection and analysis method provided by this invention also has the advantages of low analysis cost, good separation effect, strong specificity, and automated operation; thus ensuring the quality control of tacrolimus capsules. It can reduce the analysis costs for manufacturing enterprises, continuously improve the quality standards of this type of drug, thereby improving drug quality and further protecting the medication safety of the general public.

[0035] The above descriptions are merely some embodiments of the present invention. It should be noted that those skilled in the art can make other modifications and improvements without departing from the inventive concept of the present invention, and these all fall within the protection scope of the present invention.

Claims

1. A method for the determination of the dissolution profile of tacrolimus capsules, characterized in that, The determination is performed by using high performance liquid chromatograph, the chromatographic column is Prodigy Phenyl-3 phenyl column; the mobile phase is composed of acetonitrile-water-methanol-diluted phosphoric acid, the proportion of the mobile phase is 460:360:180:1; Dissolution medium: pH 6.8, pH 4.5; Preparation of test sample solution: Take tacrolimus capsules, take samples at 5 min, 10 min, 15 min, 20 min, 30 min, 45 min, 60 min, 90 min, 120 min, filter the samples through 0.45 μm microporous filter membrane, take 5 ml of the filtered solution, add 2 ml of acetonitrile, mix well, and obtain; Preparation of reference substance solution: Accurately take a proper amount of tacrolimus reference substance, dissolve and dilute to prepare a solution containing about 0.015 mg per 1 ml, take 2 ml, add dissolution medium, mix well, and obtain; Take 400 μl of test sample solution and reference substance solution respectively, inject into the liquid chromatograph respectively, record the chromatogram, and calculate the dissolution at each time point according to the peak area by external standard method.

2. The method of claim 1, wherein, The chromatographic conditions are as follows: column temperature 45 ℃; flow rate 1.0 ml / min; detection wavelength 210 nm.

3. The method of claim 2, wherein, The chromatograph is Shimadzu high performance liquid chromatograph.

4. The method of claim 2, wherein, The dissolution rotation speed of the dissolution medium is 50 rpm, 75 rpm.

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