Liquid phase detection method for dissolution rate of allopurinol tablets

Abstract

The invention discloses a method for determining the dissolution rate and the dissolution curve of purinol tablets. The dissolution rate of the allopurinol tablets is detected by high performance liquid chromatography instead of an ultraviolet spectrophotometer method used at present. According to the method, a chromatographic column is used as a silica gel bonded octadecylsilane column, an acid solution-organic phase is used as a mobile phase, and an isocratic elution mode is adopted for detection. According to the method for detecting the dissolution rate and the dissolution curve of the allopurinol tablet through the high performance liquid chromatography, detection can be easily and conveniently completed, specificity is high, accuracy is high, repeatability is good, application and linearity are achieved, and the dissolution rate of the allopurinol tablet and the dissolution amount of the dissolution curve can be efficiently and rapidly detected.

Description

Technical Field

[0001] Through exploratory research, an HPLC method for determining the dissolution rate of allopurinol tablets was established. Research on related products can refer to the proposed method to focus on the dissolution rate of the drug. Background Technology

[0002] Allopurinol, also known as xanthine oxidase inhibitor, is a chemical inhibitor of xanthine oxidase. Its chemical name is 4-hydroxypyrazolo[3,4-d]pyrimidine, with the chemical formula C5H4N4O and a molecular weight of 136.11.

[0003] Its structural formula is as follows:

[0004] Allopurinol was approved for medical use in the United States in 1996. This drug was the 40th most frequently prescribed medication in the US in 2021, with over 15 million prescriptions issued that year.

[0005] Because ultraviolet spectrophotometry offers relatively high precision within the absorbance range of 0.3–0.7, and the two-step dilution preparation of the test solution during dissolution curve detection is extremely labor-intensive, it significantly increases the workload for determining bioequivalence and assessing differences in drug performance in the in vivo environment. Therefore, developing an HPLC method for the dissolution of allopurinol tablets, enabling the study of the entire process of drug entry into liquid form, is of great significance for the quality control of pharmaceutical formulations. Summary of the Invention

[0006] This invention proposes a liquid chromatography method for the dissolution of allopurinol tablets, which can quickly, effectively, simply and conveniently complete the detection, and solves the problem of huge workload in the two-step dilution preparation of the test solution in the dissolution curve detection experiment. This enables the determination of the bioequivalence of allopurinol tablets and the evaluation of the drug's performance in the in vivo environment.

[0007] The present invention proposes a liquid chromatography method for detecting the dissolution rate of allopurinol tablets. The method employs high performance liquid chromatography (HPLC) with a silica-bonded octadecylsilane column. A certain proportion of acid solution is used as mobile phase A and an organic phase is used as mobile phase B for isocratic elution.

[0008] The liquid phase detection method for allopurinol tablet dissolution provided by the present invention includes an isocratic elution procedure of 0.125% phosphoric acid solution-methanol (90:10).

[0009] Preferably, the acid solution is a 0.125% phosphoric acid solution.

[0010] Preferably, the chromatographic column provided is a BDS Hypersil C18 (4.6 mm × 150 mm, 5 µm).

[0011] The HPLC method for dissolution determination described in this invention can be implemented according to the following method: 1) Accurately weigh 10 mg of allopurinol reference standard, place it in a 10 ml volumetric flask, add 4 ml of 0.1 mol / L sodium hydroxide solution, sonicate for 1 min to dissolve, dilute with water to the mark, shake well, accurately measure 5 ml, place it in a 50 ml volumetric flask, dilute with 0.1 NHCl solution to the mark, and shake well; 2) Dissolution conditions: Use 1000 ml of phosphate buffer (pH 6.8) as the dissolution medium, rotate at 75 rpm, and maintain a temperature of 37°C. Operate according to the procedure, and take samples after 30 minutes. Take an appropriate amount of the dissolution solution, filter it, and use the filtrate as the test solution. 3) Set the mobile phase flow rate to 0.9-1.1 mL / min, preferably 1.0 mL / min, the detection wavelength to 265-275 nm, the optimal detection wavelength to 270 nm, and the column oven temperature to 30-40 °C, with the optimal temperature to 35 °C. 4) Place approximately 50 mg of allopurinol reference standard in a 50 mL volumetric flask, add 10 mL of 0.1 N sodium hydroxide solution, shake for 10 min, dilute to the mark with water, mix well, and filter to obtain the allopurinol reference stock solution. Accurately measure 3.2 mL, 3.6 mL, 4 mL, 4.4 mL, and 4.8 mL of each solution into 200 mL volumetric flasks, dilute to the mark with the mobile phase, and mix well to obtain solutions with relative concentrations of 80%, 90%, 100%, 110%, and 120%, respectively. 5) Place the reference solution and the test solution at room temperature and take samples at 0, 2, 4, 6, 8, and 12 hours, respectively; 6) Take 10 μl of each of the above sample solutions and inject it into the liquid chromatograph to complete the determination.

[0012] High-performance liquid chromatograph: Agilent 1260 Infinity II; Column: BDS Hypersil C18 (4.6 mm × 150 mm, 5 µm); Mobile phase A: 0.125% phosphoric acid solution; Mobile phase B: Methanol; Perform isocratic elution: Flow rate: 1.0 mL / min; Column temperature: 35℃; Detection wavelength: 270nm; Injection volume: 10 μl; The invention utilizes a BDS Hypersil C18 (150 × 4.6 mm, 5 μm) chromatographic column, enabling rapid, efficient, and convenient detection of allopurinol tablet dissolution. This invention solves the problem of the enormous workload associated with the two-step dilution preparation of the test solution in dissolution curve detection experiments, ensuring the quality control of in vitro release studies of allopurinol tablets. Attached Figure Description

[0013] Figure 1 This is an HPLC chromatogram showing the system suitability of the allopurinol tablet dissolution analysis method in Example 1.

[0014] Figure 2 This is a repeatable HPLC chromatogram of the allopurinol tablet dissolution analysis method in Example 1.

[0015] Figure 3 The figure shows the linearity and range HPLC chromatogram of the allopurinol tablet dissolution analysis method in Example 1.

[0016] Figure 4 , Figure 5 The figures show the HPLC chromatograms of the reference solution and the test solution for the solution stability analysis method of allopurinol tablets in Example 1. Detailed implementation method: The following embodiments are provided to further understand the present invention, but are not limited to the scope of these embodiments.

[0017] Example 1 Instruments and conditions High-performance liquid chromatograph: Agilent 1260 Infinity II; Column: BDS Hypersil C18 (4.6 mm × 150 mm, 5 µm); Mobile phase: 0.125% phosphoric acid solution - methanol (90:10) Flow rate: 1.0 mL / min; Column temperature: 35℃; Detection wavelength: 270nm; Injection volume: 10 μl; The HPLC detection steps for the dissolution of allopurinol tablets are as follows: Accurately weigh 10 mg of allopurinol reference standard and place it in a 10 ml volumetric flask. Add 4 ml of 0.1 mol / L sodium hydroxide solution, sonicate for 1 min to dissolve, then dilute with water to the mark and shake well. Accurately measure 5 ml of the solution and place it in a 50 ml volumetric flask. Dilute with 0.1 NHCl solution to the mark and shake well.

[0018] Dissolution conditions: 1000 ml of phosphate buffer (pH 6.8) was used as the dissolution medium; the rotation speed was 75 rpm; and the temperature was 37°C. The procedure was followed, and samples were taken after 30 minutes. An appropriate amount of the dissolution solution was taken, filtered, and the filtrate was used as the test solution. Inject 10 µl of the above solution into the liquid chromatograph and record the chromatogram.

[0019] System suitability test results:

[0020] Repeatability test results:

[0021] Linearity and Range Experiment Results:

[0022] Solution stability test results: 1. Allopurinol reference solution

[0023] 2. Sample solution

[0024] In summary, this invention enables simple and convenient detection, with strong specificity, high accuracy, good repeatability, applicability, and linearity. It can efficiently and rapidly detect the dissolution rate and dissolution amount of allopurinol tablets based on the dissolution curve.

Claims

1. A liquid chromatography method for detecting the dissolution rate of allopurinol tablets, comprising high performance liquid chromatography, using a silica-bonded octadecylsilane column, and using an acid solution-organic phase as the mobile phase for isocratic elution.

2. The liquid chromatography method for detecting the dissolution of allopurinol tablets according to claim 1, wherein the chromatographic column is selected from one of the following silica-bonded octadecylsilane columns: Promosil C18, BDS Hypersil C18.

3. The liquid phase detection method for the dissolution of allopurinol tablets according to claim 1, wherein the acid solution is selected from one of the following proportions: 0.1125% phosphoric acid solution, 0.125% phosphoric acid solution, or 0.1375% phosphoric acid solution.

4. The liquid chromatography method for detecting the dissolution rate of allopurinol tablets according to claim 1, wherein the elution process is isocratic elution: 0.125% phosphoric acid solution - methanol (90:10).

5. The liquid chromatography method for detecting the dissolution rate of allopurinol tablets according to claim 1, wherein the chromatographic column is preferably 150 mm in length, 4.6 mm in diameter, and has a packing particle size of 5 µm.

6. In the liquid phase detection method for the dissolution of allopurinol tablets according to claims 1 and 3, the required acid solution is preferably a 0.125% phosphoric acid solution.

7. The liquid chromatography method for detecting the dissolution rate of allopurinol tablets according to claims 2 and 5, wherein the chromatographic column is preferably a BDSH Hypersil C18.

8. The liquid chromatography method for detecting the dissolution rate of allopurinol tablets according to claim 1 includes the following steps: ① setting the flow rate to 0.9~1.1 ml / min; ② setting the detection wavelength to 265-275 nm; ③ setting the column temperature to 30~40℃.

9. The liquid chromatography method for detecting the dissolution rate of allopurinol tablets according to claim 8, wherein the chromatographic conditions are as follows: ① the flow rate is preferably 1.0 ml / min; ② the wavelength is preferably 270 nm; ③ the column temperature is preferably 35 °C.

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