A method for detecting iodine content in dioxymatin bulk drug and a quality control method for iodine content

Abstract

The application discloses a method for detecting iodine content in diltiaxin bulk drug and a quality control method of the iodine content, and belongs to the technical field of drug detection and quality control, and comprises the following steps: preparing a control solution of potassium iodide; dissolving the to-be-detected substance in a hydrazine solution after combustion to prepare a test solution; performing liquid chromatography detection on the control solution and the test solution to obtain corresponding chromatograms; and calculating the iodine content in the to-be-detected substance according to the concentration and peak area of the control solution and the concentration and peak area of the test solution in the chromatograms. The application can quantitatively detect the residual iodine content based on the liquid chromatography detection method, the analysis result is reliable, and the application has the advantages of high detection precision, simple operation steps, high detection efficiency and the like.

Description

TECHNICAL FIELD

[0001] The present application is a method for detecting the iodine content in diosmin bulk drug and a quality control method for the iodine content, specifically relates to a method for detecting the iodine content in diosmin bulk drug by high performance liquid chromatography, and a quality control method for the iodine content in diosmin bulk drug by high performance liquid chromatography, and belongs to the technical field of drug detection and quality control. BACKGROUND

[0002] Bulk drug is an effective component for producing various preparations, and is usually prepared by chemical synthesis, plant extraction or biotechnology to obtain powders, crystals, extracts and the like with medicinal properties, which are then processed into pharmaceutical preparations to obtain drugs for clinical use. Therefore, the quality of bulk drug determines the quality of preparations. In order to widely use bulk drug and its related preparations, it is necessary to establish a quality control method, especially to strengthen the control of toxic components in drugs and the qualitative and quantitative research of impurities in drugs, so as to ensure the effectiveness and safety of drugs.

[0003] Diosmin is a drug that can enhance venous tension, reduce vascular fragility and abnormal permeability, promote lymphatic return and improve microcirculation. The current common synthesis method of diosmin is to dissolve hesperidin in pyridine, add iodine to undergo iodination reaction, then dehydrogenate to obtain crude product, and then purify by water elution to obtain finished bulk drug. Therefore, the content of residual iodine in the finished bulk drug needs to be detected to realize the quality control of diosmin bulk drug.

[0004] In the prior art, the patent for invention with publication number CN104730068A discloses a method for determining the iodine residue in diosmin bulk drug, which is determined by potential titration after using a selective membrane electrode potential specifically for iodine ions, wherein the reference electrode is Ag / AgCl (silver / silver chloride), the internal electrolyte is LiCl saturated in ethanol, the external electrolyte is KNO3, a saturated aqueous solution, and the electrolyte is a 2M KNO3 solution in 0.1N nitric acid. However, this method can only be used for qualitative detection, and cannot be used for quantitative detection. In addition, there are problems such as high operation difficulty, poor detection accuracy and reproducibility, etc. during detection, so it is not conducive to realizing the quality control of diosmin bulk drug.

[0005] In addition, the prior art also discloses that ion chromatography is used to detect iodine residues in diosmin raw materials, specifically, after online combustion pretreatment of a detection sample, absorption by an alkali solution, separation of iodine ions by ion chromatography, and detection by an amperometric detector, wherein an analysis column is SH-G-1+SH-AC-17; a mobile phase is 10.0 mM KOH; a flow rate is 1.0 mL / min; a column temperature is 35 DEG C; a detection mode is direct current amperometric detection; a sample injection volume is 25 mu L; and pretreatment is that the sample is subjected to online combustion pretreatment, and the absorption liquid (S2O3 2- ) is filtered through a 0.22 mu m filter membrane for sample analysis. However, in the sample preparation process, the method is prone to introduce interference, causing unstable baseline and high noise during detection, which interferes with the detection result, and then causes a high detection limit, and when the iodine content is low, the iodine content cannot be accurately detected, and since the sample needs to be separated by ion chromatography before detection, the detection time is long, and therefore, the method is not conducive to quality control of diosmin raw materials. SUMMARY

[0006] The purpose of the present application is to provide a method for detecting iodine content in diosmin raw materials and a quality control method for iodine content, which can quantitatively detect residual iodine content based on liquid chromatography detection, and the analysis result is reliable, and the quality control of iodine content in diosmin raw materials can be realized, which has important application value for quality control of raw materials.

[0007] The present application is realized by the following technical scheme: a method for detecting iodine content in diosmin raw materials, comprising the following steps:

[0008] S1. preparing a potassium iodide control solution;

[0009] S2. dissolving the combustion sample in a hydrazine solution to obtain a test sample solution;

[0010] S3. liquid chromatography detection of the control solution and the test sample solution to obtain corresponding chromatograms;

[0011] S4. calculating the iodine content in the test sample according to the concentration and peak area of the control solution and the peak area of the test sample solution in the chromatogram.

[0012] In step S1, 1-20.0 mg of potassium iodide is dissolved in 50-100 ml of pure water to obtain an aqueous solution of potassium iodide, then 1-5 ml of the aqueous solution is taken and diluted with 20-50 ml of a hydrazine solution with a concentration of 0.1-0.5 g / L, and then 1-5 ml of the diluted solution is taken and 5-15 ml of a mobile phase is added.

[0013] In the step S2, 0.100-0.300 g of the test substance is wrapped in filter paper and placed on a sample holder, 20-50 ml of a hydrazine solution is injected into a flask, oxygen is introduced into the flask, the filter paper is ignited, and after the combustion is completed, the combustion product is immediately stirred to completely dissolve, 1-5 ml of the dissolved solution is weighed, and 5-15 ml of a mobile phase is added to obtain the solution.

[0014] In the step S3, 50 μl of the control solution and the test solution are respectively injected into a liquid chromatograph, a C18 chromatographic column is used, methanol and 3 mmol / L tetrabutylammonium bromide in a volume ratio of 30:70 are used as the mobile phase, isocratic elution is carried out for 10-20 min, the flow rate is controlled at 0.8-1.2 ml / min, the column temperature is 25-35 °C, and the detection wavelength is 220-240 nm.

[0015] In the step S4, the calculation formula of the iodine content is shown in the following formula (1):

[0016] (1)

[0017] In the above formula (1), A test is the peak area of the test sample, C control is the concentration of the control sample, A control is the peak area of the control sample, C test is the concentration of the test sample, 127 is the molar mass of iodine ions, and 166 is the molar mass of potassium iodide.

[0018] The application also provides a quality control method for the iodine content in diosmin bulk drug, which can be used to determine the iodine content in diosmin bulk drug by the above method, and the effectiveness and safety of diosmin bulk drug can be ensured by controlling the iodine content in diosmin bulk drug to be ≤0.1%.

[0019] Compared with the prior art, the application has the following advantages and beneficial effects:

[0020] (1) The application uses a hydrazine solution to dissolve the test substance after combustion to prepare a test solution, dilutes potassium iodide with a hydrazine solution to prepare a control solution, and uses high-performance liquid chromatography to detect the iodine content in diosmin bulk drug, which can avoid the influence of impurities on the detection results, realize accurate detection of the iodine content, and the detection limit reaches 0.0061 ug / ml.

[0021] (2) The application is based on high-performance liquid chromatography and specific detection conditions, can quickly and accurately obtain the characteristic spectrum of the test substance, and can calculate the iodine content in the test substance through the chromatogram of the control solution, which not only realizes quantitative detection of the iodine content, but also has the advantages of high detection precision, simple operation steps, and high detection efficiency compared with the existing amperometric detection method.

[0022] (3) The method can provide a detection standard for quality control of iodine content in dioxymatin raw medicine, can effectively control the impurity iodine content in dioxymatin raw medicine to be not more than 0.1%, thereby guaranteeing the effectiveness and safety of dioxymatin raw medicine. BRIEF DESCRIPTION OF DRAWINGS

[0023] Figure 1 The chromatogram of the test sample solution in Example 1 of the present application.

[0024] Figure 2 The chromatogram when the blank sample is used in the specificity determination of the present application.

[0025] Figure 3 The chromatogram when the blank solvent sample is used in the specificity determination of the present application.

[0026] Figure 4 The chromatogram when the control substance stock solution sample is used in the specificity determination of the present application.

[0027] Figure 5 The chromatogram when the control solution sample is used in the specificity determination of the present application.

[0028] Figure 6 The chromatogram when the test sample solution sample is used in the specificity determination of the present application (the same as Figure 1 ).

[0029] Figure 7 The chromatogram when the limit of quantification determination of the present application is determined (one).

[0030] Figure 8 The chromatogram when the limit of quantification determination of the present application is determined (two).

[0031] Figure 9 The chromatogram when the limit of quantification determination of the present application is determined (three).

[0032] Figure 10 The chromatogram when the limit of quantification determination of the present application is determined (four).

[0033] Figure 11 The chromatogram when the limit of quantification determination of the present application is determined (five).

[0034] Figure 12 The chromatogram when the limit of quantification determination of the present application is determined (six).

[0035] Figure 13 The chromatogram when the limit of detection determination of the present application is determined. DETAILED DESCRIPTION

[0036] The present application will be further described in detail below in combination with examples, but the embodiments of the present application are not limited thereto.

[0037] The dioxadromine bulk drug used in the following examples is from Chengdu Yizhong Biopharmaceutical Co., Ltd., with batch number 4003a-20230401.

[0038] Example 1: Determination of the content of iodine in dioxadromine bulk drug

[0039] The method for determining the content of iodine in dioxadromine bulk drug in this example is as follows:

[0040] Preparation of the reference solution: precisely weigh 12.8 mg of potassium iodide, dissolve and dilute to 100.0 mL with pure water; precisely take 1.0 mL of the solution, dilute to 50.0 mL with a hydrazine solution, precisely take 1.0 mL, and add to a 10 mL volumetric flask, and add the mobile phase (methanol: 3 mmol / L tetrabutylammonium bromide = 30:70) to the calibration mark;

[0041] Preparation of the test solution: wrap 0.100 g of the test substance in filter paper, place it in a sample holder, inject 50 mL of a hydrazine solution into a flask, introduce oxygen into the flask for 10 minutes, ignite the filter paper, immediately after combustion, stir to completely dissolve the combustion product, and then continue stirring for 1 hour; precisely take 1.0 mL of the solution, dilute to 10 mL with the mobile phase (methanol: 3 mmol / L tetrabutylammonium bromide = 30:70), and obtain the test solution;

[0042] Determination: determine the above reference solution and test solution according to the following chromatographic conditions, and record the chromatogram as shown in Figure 1 , according to the concentration, peak area of the reference solution, and the concentration, peak area of the test solution, calculate the content of iodine in the test sample by external standard method.

[0043] Specifically, the calculation formula of the content of iodine is shown in formula (1) as follows:

[0044] (1)

[0045] Wherein, Atest: test peak area; Cref: reference concentration; Aref: reference peak area; Ctest: test concentration; 127: molar mass of iodine ion; 166: molar mass of potassium iodide.

[0046] When determining, the chromatographic conditions used are as follows:

[0047] Chromatographic column: waters ODS C18 5um, 4.6mm x 250mm;

[0048] Injection volume: 50ul;

[0049] Mobile phase: methanol: 3 mmol / L tetrabutylammonium bromide = 30:70, isocratic elution;

[0050] Elution time: 15 min;

[0051] Column temperature: 30 °C;

[0052] Flow rate: 1.0 ml / min;

[0053] Detection wavelength: 223 nm.

[0054] In this example, according to the concentration of the reference solution (0.256 ug / m1) and the peak area 59.37 and the peak area of the test sample 29.26, the iodine content in the raw material of Dioxybenzone was calculated by external standard method as 0.048%, not more than 0.1%, meeting the quality control requirements of impurity iodine content in Dioxybenzone raw material.

[0055] Example 2: Methodology verification

[0056] According to the analysis method verification guidelines of the People's Republic of China Pharmacopoeia 2020 edition (9101) and ICH analysis method verification guidelines, the detection method of example 1 was verified, and the process and results were as follows:

[0057] The solution was prepared as follows:

[0058] Preparation of reference stock solution: accurately weigh potassium iodide 12.8 mg, dissolve and dilute to 100.0 mL with pure water. Accurately take 1.0 mL of the solution, dilute to 50.0 mL with hydrazine solution to obtain the stock solution.

[0059] Preparation of reference solution: accurately take 1.0 ml of the stock solution into a 10 ml volumetric flask, add mobile phase to the mark.

[0060] Preparation of test sample solution: wrap 0.100 g of the test substance in filter paper, place it in a sample holder, inject 50 mL of 0.2 g / L hydrazine solution into the flask, pass oxygen through the flask for 10 minutes, ignite the filter paper, immediately stir after burning to completely dissolve the combustion product, then continue stirring for one hour; accurately take 1.0 ml of the solution, dilute with mobile phase to 10 ml, and obtain the test sample solution.

[0061] Preparation of blank solvent: take filter paper of the same size as the test sample, place it in a sample holder, inject 50 mL of 0.2 g / L hydrazine solution into the flask, pass oxygen through the flask for 10 minutes, ignite the filter paper, immediately stir after burning to completely dissolve the combustion product, then continue stirring for one hour. Accurately take 1.0 ml of the solution, dilute with mobile phase to 10 ml, and obtain the blank solvent.

[0062] (I) System suitability

[0063] Take the control solution, the method of Example 1 was determined, 6 needles. The results are shown in Table 1.

[0064] (II) Specificity

[0065] The above three kinds of solution and blank solvent, respectively, according to the method of Example 1 was determined, recorded the corresponding chromatogram, see Figures 2 to 6 , Figures 2 to 6 In turn, blank, blank solvent, control solution, control solution, test solution. The results are shown in Table 1.

[0066] (1) detection limit and limit of quantification

[0067] Take the control solution of 10% solution, investigate the signal to noise ratio, greater than 10 can be used as the limit of quantification concentration, dilute 3 times the limit of quantification concentration sample, investigate the signal to noise ratio, signal to noise ratio greater than 3 can be used as the detection limit concentration, according to the method of Example 1 was determined.

[0068] The limit of quantification concentration solution was continuously sampled 6 needles, recorded the corresponding chromatogram, see Figures 7 to 12 ; the detection limit concentration sample 1 needle, recorded the corresponding chromatogram, see Figure 13 . The results are shown in Table 1.

[0069] (2) linearity and range

[0070] Precise control of the control solution 1.0, 5.0, 8.0, 10.0, 15.0 ml, respectively, in 100 ml flask, add mobile phase dilution to the mark, according to the method of Example 1 was determined. The results are shown in Table 1.

[0071] (3) precision

[0072] (3.1) repeatability

[0073] The same test sample, preparation of 6, according to the method of Example 1 was determined. The results are shown in Table 1.

[0074] (3.2) intermediate precision

[0075] The same sample, different personnel with different instruments and equipment (Agilent / Thermo), on different dates, according to the method of Example 1 was determined. The results are shown in Table 1.

[0076] (4) accuracy

[0077] Precise control of the control solution 0.5, 1.0, 1.5 ml, respectively, in 20 ml flask, add mobile phase dilution to the mark, shake, the same method preparation 3. As the concentration of accuracy solution, according to the method of Example 1 was determined. The results are shown in Table 1.

[0078] (5) Solution stability

[0079] The control solution and the test solution were placed at room temperature, and samples were taken at 0 and 24 h, and then determined according to the method of Example 1. The determination results are shown in Table 1.

[0080] (6) Durability

[0081] The control solution and the test solution were determined according to the method of Example 1 at different flow rates (the flow rates were 0.8 ml / min, 1.0 ml / min and 1.2 ml / min, respectively) and column temperatures (the column temperatures were 25℃, 30℃ and 35℃, respectively) for the content of diosmin iodine. The determination results are shown in Table 1.

[0082] Table 1 Summary of determination method verification for the content of diosmin iodine

[0083]

[0084] The above description is only the preferred embodiments of the present application, and does not limit the present application in any form. Any simple modification or equivalent change made according to the technical essence of the present application to the above embodiments falls within the protection scope of the present application.

Claims

1. A method for detecting the iodine content in diltiaxin bulk drug substance, characterized by: The method comprises the following steps: S1. Weigh 1-20.0 mg of potassium iodide and dissolve it in 50-100 ml of pure water to obtain a potassium iodide aqueous solution, then take 1-5 ml of the aqueous solution, add 20-50 ml of a dilute hydrazine solution with a concentration of 0.1-0.5 g / L, take 1-5 ml of the diluted solution, and add 5-15 ml of a mobile phase to prepare a reference solution of potassium iodide; S2. Wrap 0.100-0.300 g of the test substance in filter paper, place it on a sample holder, inject 20-50 ml of a hydrazine solution into a flask, introduce oxygen into the flask, ignite the filter paper, immediately stir after combustion to completely dissolve the combustion product, and continue stirring for 0.5-1.5 hours, then take 1-5 ml of the dissolved solution, add 5-15 ml of a mobile phase to prepare a test solution; S3. Take 50 μl of the reference solution and the test solution respectively, inject them into a liquid chromatograph, use methanol and 3 mmol / L tetrabutylammonium bromide with a volume ratio of 30:70 as the mobile phase, perform isocratic elution for 10-20 min, control the flow rate at 0.8-1.2 ml / min, the column temperature at 25-35℃, and perform liquid chromatography detection at a detection wavelength of 220-240 nm to obtain the corresponding chromatogram; S4. According to the concentration and peak area of the reference solution and the concentration and peak area of the test solution in the chromatogram, the content of iodine in the test substance is calculated, and the detection limit concentration of iodine in the test substance is 0.0061 ug / ml.

2. The method of claim 1, wherein: In the step S4, the calculation formula of the iodine content is shown in the following formula (1): （1） In the above formula (1), Atest: test peak area; Cref: reference concentration; Aref: reference peak area; Ctest: test concentration; 127: molar mass of iodine ion; 166: molar mass of potassium iodide.

3. A method for quality control of the iodine content in dioxadimine bulk drug substance, characterized by: The iodine content in the diosmin bulk drug is determined by the method of any one of claims 1 or 2, and the iodine content in the diosmin bulk drug is controlled to be ≤0.1%.

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