A method for detecting 5-hydroxymethylfurfural in the compatibility experiment of hydrocortisone sodium succinate for injection and glucose injection by HPLC

Abstract

The application discloses a method for detecting 5-hydroxymethylfurfural in a compatibility experiment of hydrocortisone sodium succinate for injection and glucose injection by HPLC. The 5-hydroxymethylfurfural is a degradation impurity of glucose. The detection method comprises the following steps: (1) preparation of a control sample solution; (2) preparation of a test sample solution; (3) chromatographic conditions; a mobile phase A is 8mmol / L potassium dihydrogen phosphate solution (pH value is adjusted to 5.0-5.1 by using 8mmol / L potassium hydrogen phosphate solution, and the solution is prepared freshly before use); a mobile phase B is methanol, gradient elution is carried out; a detection wavelength is 284nm; a column temperature is 40 DEG C; a flow rate is 1.4-1.6mL / min; and a chromatographic column is C 18 ; the application can effectively detect the content of 5-hydroxymethylfurfural, and has strong specificity and high sensitivity.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical quality control, specifically relating to an HPLC method for detecting 5-hydroxymethylfurfural in a compatibility test of hydrocortisone sodium succinate for injection and glucose injection. Background Technology

[0002] Hydrocortisone sodium succinate for injection is an adrenocortical hormone. It possesses various pharmacological effects, including anti-inflammatory, anti-allergic, and immunosuppressive properties. It improves blood flow by reducing blood viscosity. It is primarily used for adrenocortical insufficiency and pituitary hypofunction, and is also used for allergic and inflammatory diseases.

[0003] In a compatibility experiment with glucose injection, sodium hydrocortisone succinate for injection may promote the production of 5-hydroxymethylfurfural.

[0004] 5-hydroxymethylfurfural may be generated during the sample compatibility stability process, and compatibility at different concentrations needs to be controlled. This is of great practical significance for improving the quality control of injectable hydrocortisone sodium succinate. Summary of the Invention

[0005] The purpose of this invention is to solve the above problems by providing an HPLC method for separating and detecting 5-hydroxymethylfurfural in a compatibility experiment of injectable hydrocortisone sodium succinate and glucose injection. This method can simultaneously detect the content of impurity 1 and impurity 2 in injectable hydrocortisone sodium succinate, and has the characteristics of high specificity, accurate localization, high sensitivity and stability.

[0006] The objective and technical solution of this invention is as follows: In the compatibility experiment of hydrocortisone sodium succinate for injection and glucose injection, the separation and detection of 5-hydroxymethylfurfural is performed using high-performance liquid chromatography (HPLC). The steps include:

[0007] (1) Preparation of the test solution:

[0008] Take an appropriate amount of this product, hydrocortisone sodium succinate for injection, add an appropriate amount of compatibility solution (5% glucose injection, glucose sodium chloride injection), dissolve, and shake well to obtain the product. (Contains hydrocortisone 50mg / ml, 2mg / ml, 0.1mg / ml).

[0009] (2) Preparation of reference solution:

[0010] Take an appropriate amount of 5-hydroxymethylfurfural reference standard, dissolve and dilute it with water to prepare a solution containing approximately 10 μg per 1 ml.

[0011] (3) Testing:

[0012] Inject 20 μL each of the test solution and the reference solution into the high-performance liquid chromatograph (HPLC) for gradient elution. The chromatographic conditions of the HPLC are as follows:

[0013] Mobile phase A: 8 mmol / L potassium dihydrogen phosphate solution (adjusted to pH 5.0-5.1 with 8 mmol / L potassium hydrogen phosphate solution, freshly prepared before use); Mobile phase B: methanol; Elute according to a gradient elution program.

[0014] Flow rate: 1.4–1.6 mL / min; preferably 1.5 mL / min.

[0015] Column temperature: 40℃.

[0016] Column: C 18 Column; preferably Hypersil BDS-C 18 The dimensions are 4.6mm × 250mm, and the micrometer size is 5μm.

[0017] Detection wavelength: 284nm.

[0018] The specific gradient elution conditions are as follows:

[0019] The volume percentage of mobile phase B was 10% from 0 to 5 min. From 5.01 to 10 min, the volume percentage of mobile phase B increased uniformly from 10% at 5.01 min to 45% at 10 min. From 10.01 to 30 min, the volume percentage of mobile phase B remained at 45%. From 30.01 to 31 min, the volume percentage of mobile phase B decreased from 45% at 30.01 min to 10% at 31 min. From 31.01 to 35 min, the volume percentage of mobile phase B remained at 10%.

[0020] The positive effects of this invention are as follows: This invention uses high performance liquid chromatography, which can effectively detect the content of 5-hydroxymethylfurfural. This method has high specificity, high sensitivity, and simple operation, and can effectively improve the quality control system of injectable hydrocortisone sodium succinate, thereby ensuring the safety and effectiveness of the product. Attached Figure Description

[0021] Figure 1 This is the chromatogram of the blank solution in the experimental example;

[0022] Figure 2 This is the chromatogram of the control solution in the experimental example;

[0023] Figure 3 The chromatogram of the test solution is shown in the example. Detailed Implementation

[0024] Test case

[0025] The content of 5-hydroxymethylfurfural in a compatibility test of hydrocortisone sodium succinate for injection and glucose injection was determined by high performance liquid chromatography.

[0026] Reagents: potassium dihydrogen phosphate, dipotassium hydrogen phosphate, methanol, water.

[0027] Test reagent: 5-hydroxymethylfurfural reference standard.

[0028] Instruments: High-performance liquid chromatograph: Shimadzu LC-2030; Hypersil BDS-C 18 , 4.6mm×250mm, 5μm.

[0029] Chromatographic conditions: Mobile phase A: 8 mmol / L potassium dihydrogen phosphate solution (pH adjusted to 5.0–5.1 with 8 mmol / L dipotassium hydrogen phosphate solution, freshly prepared before use); Mobile phase B: methanol; Elution using a gradient elution program; Flow rate: 1.5 mL / min; Column temperature: 40 °C; Detection wavelength: 284 nm; Injection volume: 20 μL.

[0030] Experimental Procedure: Take an appropriate amount of 5-hydroxymethylfurfural reference standard, dissolve and dilute it with water to prepare a solution containing approximately 10 μg per ml, as the reference solution. Accurately measure 20 μl each of the above blank solvent and system adaptability solution, analyze them according to the above chromatographic conditions, record the chromatograms, and see the results below. Figure 1 and Figure 2 Chromatography Figure 2 The chromatographic peak at a retention time of 4.440 min is that of 5-hydroxymethylfurfural. (From chromatogram) Figure 2 The results show that the blank solvent does not interfere with the elution of 5-hydroxymethylfurfural peaks, and the plate number of 5-hydroxymethylfurfural is 15891, indicating a good peak shape.

[0031] Example

[0032] The instruments and chromatographic conditions were the same as in the experimental example. Reagents: potassium dihydrogen phosphate, dipotassium hydrogen phosphate, methanol, and water. Chemicals: sodium hydrocortisone for injection and glucose injection.

[0033] Experimental steps:

[0034] Take an appropriate amount of this product, hydrocortisone sodium succinate for injection, add an appropriate amount of compatibility solution (5% glucose injection, glucose sodium chloride injection), dissolve, and shake well to prepare the test solution.

[0035] Accurately measure 20 μL of the above test solution, analyze it under the above chromatographic conditions, record the chromatogram, and the results are shown in the figure. Figure 3 .

[0036] Combination Figure 1 , Figure 2 , Figure 3Analysis shows that the method of this invention exhibits a good peak shape for 5-hydroxymethylfurfural and meets the required sensitivity, making it accurate for the quantitative detection of 5-hydroxymethylfurfural in the compatibility experiment of hydrocortisone sodium succinate and glucose injection.

Claims

1. A method for separating and detecting 5-hydroxymethylfurfural in a compatibility test of hydrocortisone sodium succinate for injection and glucose injection using HPLC, the detection method comprising the following steps: (1) Preparation of the test solution: Take an appropriate amount of this product, hydrocortisone sodium succinate for injection, add an appropriate amount of compatibility solution (5% glucose injection, glucose sodium chloride injection), dissolve, and shake well to obtain the product. (Contains hydrocortisone 50mg / ml, 2mg / ml, 0.1mg / ml); (2) Preparation of reference solution: Take an appropriate amount of 5-hydroxymethylfurfural reference standard, dissolve and dilute it with water to prepare a solution containing approximately 10 μg per 1 ml; (3) Testing: Inject 20 μl each of the test solution and the reference solution into the high-performance liquid chromatograph and perform gradient elution. The chromatographic conditions of the high-performance liquid chromatograph are as follows: Mobile phase A: 8 mmol / L potassium dihydrogen phosphate solution (pH adjusted to 5.0–5.1 with 8 mmol / L dipotassium hydrogen phosphate solution, freshly prepared before use); Mobile phase B: methanol; Elution using a gradient elution program: flow rate: 1.4–1.6 mL / min, column temperature: 40 °C, column: C 18 Column; Detection wavelength: 284nm; The specific gradient elution conditions are as follows: The volume percentage of mobile phase B was 10% from 0 to 5 min. From 5.01 to 10 min, the volume percentage of mobile phase B increased uniformly from 10% at 5.01 min to 45% at 10 min. From 10.01 to 30 min, the volume percentage of mobile phase B remained at 45%. From 30.01 to 31 min, the volume percentage of mobile phase B decreased from 45% at 30.01 min to 10% at 31 min. From 31.01 to 35 min, the volume percentage of mobile phase B remained at 10%.

2. The method for separating and detecting 5-hydroxymethylfurfural in the compatibility test of hydrocortisone sodium succinate for injection and glucose injection according to claim 1, characterized in that: The C 18 The column is Hypersil BDS-C 18 The dimensions are 4.6mm × 250mm, and the micrometer size is 5μm.

3. The method for separating and detecting 5-hydroxymethylfurfural in the compatibility test of hydrocortisone sodium succinate for injection and glucose injection according to claim 1, characterized in that: The mobile phase A is an 8 mmol / L potassium dihydrogen phosphate solution (the pH is adjusted to 5.0-5.1 with an 8 mmol / L potassium hydrogen phosphate solution and it is freshly prepared before use), and the mobile phase B is methanol.

4. The method for separating and detecting 5-hydroxymethylfurfural in the compatibility test of hydrocortisone sodium succinate for injection and glucose injection by HPLC according to claim 1, characterized in that: The column temperature was 40℃, the flow rate was 1.5mL / min, and the detection wavelength was 28472 nm.

5. The method for separating and detecting 5-hydroxymethylfurfural in the compatibility test of hydrocortisone sodium succinate for injection and glucose injection according to claim 1, characterized in that: The diluent is water.

6. The method for separating and detecting 5-hydroxymethylfurfural in a compatibility test of hydrocortisone sodium succinate for injection and glucose injection according to claim 1, characterized in that: The injection volume was 20 μl.

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