阅读说明：本技术 一种盐酸贝尼地平片中的杂质检测方法 (Method for detecting impurities in benidipine hydrochloride tablets ) 是由 王熙红 曲晓华 林丽丽 杨淑萍 于 2019-10-14 设计创作，主要内容包括：本发明涉及一种盐酸贝尼地平片中的杂质检测方法,解决了国内没有盐酸贝尼地平片中的杂质检测方法没有统一的标准方法的技术问题。本发明提供一种盐酸贝尼地平片中的杂质检测方法,包括以下步骤：供试品溶液的配制：以高效液相色谱法分别测定步骤(3)制得的系统适用性溶液、步骤(2)制得的对照品溶液和步骤(1)制得的供试品溶液试验；本发明可广泛应用于制药技术领域。(The invention relates to a method for detecting impurities in benidipine hydrochloride tablets, which solves the technical problem that no unified standard method exists in the domestic method for detecting the impurities in benidipine hydrochloride tablets. The invention provides a method for detecting impurities in benidipine hydrochloride tablets, which comprises the following steps: preparing a test solution: respectively measuring the system applicability solution prepared in the step (3), the reference substance solution prepared in the step (2) and the test substance solution prepared in the step (1) by using a high performance liquid chromatography; the invention can be widely applied to the technical field of pharmacy.)

1. A method for detecting impurities in benidipine hydrochloride tablets is characterized by adopting liquid chromatography for detection and comprising the following steps:

(1) preparing a test solution: grinding benidipine hydrochloride tablets to powder, precisely weighing an appropriate amount of fine powder (about 25mg equivalent to benidipine hydrochloride), putting the fine powder into a 50ml measuring flask, adding an appropriate amount of methanol-diluted phosphoric acid mixed solution, shaking up, diluting the mixed solution to a scale, shaking up, filtering, and taking a subsequent filtrate as a sample solution;

(2) preparing a reference substance solution: taking a proper amount of benidipine hydrochloride reference substance, precisely weighing, dissolving by using the mixed solution in the step (1), and quantitatively diluting to prepare a solution containing 5 mu g of benidipine hydrochloride in each 1ml as a reference substance solution;

(3) preparation of system applicability solution: taking a proper amount of benidipine hydrochloride reference substances and isomer reference substances, precisely weighing, dissolving by using the mixed solution, and quantitatively diluting to prepare a solution containing 0.5mg of benidipine hydrochloride and 5 mu g of isomer per 1ml, wherein the solution is used as a system applicability solution;

(4) detection liquid chromatography method and conditions:

respectively measuring the system applicability solution prepared in the step (3), the reference substance solution prepared in the step (2) and the test substance solution prepared in the step (1) by using a high performance liquid chromatography;

liquid chromatography conditions: chromatographic column using octadecylsilane chemically bonded silica as filler; using 0.05mol/L potassium dihydrogen phosphate solution (pH is adjusted to 3.0 by phosphoric acid) -methanol-tetrahydrofuran as mobile phase; detecting wavelength at 237nm, column temperature at 25 deg.C, adjusting flow rate to make peak retention time of benidipine hydrochloride about 20min, collecting system applicability solution 10 μ l, injecting into liquid chromatograph, and recording chromatogram until retention time of main component is 2 times;

the separation degree of the benidipine hydrochloride peak and the isomer peak is required to meet the requirement, and the separation degree is calculated by the peak area according to an external standard method;

(5) detection indexes are as follows: if an impurity peak exists in the chromatogram of the test solution, the peak area of the dehydrogenation derivative peak with the relative retention time of about 0.75 (corrected by multiplying by 1.6) is not more than 0.5 times (0.5%) of the main peak area of the control solution except the solvent peak, the peak area of other single impurities is not more than 0.2 times (0.2%) of the main peak area of the control solution, and the sum of the peak areas of the impurities is not more than 1.0% of the main peak area of the control solution.

2. The method of claim 1, wherein the chromatography column is an Agilent Pursuit, 4.6 x 100mm, 3 μm or equivalent performance chromatography column.

3. The method for determining impurities in benidipine hydrochloride tablets according to claim 1, wherein in the step (1), the concentration of the dilute phosphoric acid is 0.2%, and the volume ratio of the methanol to the dilute phosphoric acid is 1: 1.

4. The method for determining impurities in a benidipine hydrochloride tablet according to claim 1, wherein in the step (1), the shaking-up method is mechanical shaking for 15 min.

5. The method for detecting impurities in benidipine hydrochloride tablets according to claim 1, wherein in the step (4), the volume ratio of 0.05mol/L potassium dihydrogen phosphate solution (pH adjusted to 3.0 with phosphoric acid) in the mobile phase to methanol-tetrahydrofuran is 60:20: 20.

Technical Field

The invention relates to the technical field of pharmacy, in particular to a method for detecting impurities in benidipine hydrochloride tablets.

Background

Benidipine hydrochloride belongs to dihydropyridine calcium ion antagonists, and expands blood vessels by binding to dihydropyridine receptors that block cell membrane potential-dependent calcium ion channels, and preventing calcium ion influx into cells. Benidipine hydrochloride is widely used as a therapeutic drug for hypertension, renal essential hypertension, angina pectoris, and the like because it is safe and effective.

Benidipine hydrochloride is a dihydropyridine calcium antagonist and is usually a mixture of alpha and beta diastereomers obtained by synthetic reaction, and the alpha and beta diastereomers can be separated by recrystallization.

The pharmaceutical benidipine hydrochloride is an alpha-isomer, and is a mixture of (R, R) and (S, S) isomers (the ratio of the two is 1: 1). The chemical name is as follows: 3- [3(RS) -1-benzylpiperidin-3-yl ] 5-methyl (4RS) -2, 6-dimethyl-4- (3-nitrophenyl) -1, 4-dihydro-3, 5-dicarboxylic acid ester hydrochloride.

Pharmacological research shows that benidipine and Ca²⁺The combination of the specific parts of the channel has higher affinity than other dihydropyridine calcium ion antagonists, the dissociation of benidipine and the combination sites is slower, and the action duration is longer. Benidipine is reported to inhibit KCI-induced vasoconstriction even after being eluted for a long time. In addition, both basic research and clinical research show that benidipine has the same or better effect than the current dihydropyridine calcium antagonist, has the advantages of high tolerance and low adverse reaction rate, and is a safe and effective calcium ion antagonist. Therefore, benidipine is a safe and efficient first-choice medicine for treating hypertension, and has very wide application prospect and very great market potential.

In the prior art, the synthesis method of benidipine hydrochloride generally has the defects of low yield, more byproducts and difficult product purification, and the prepared benidipine hydrochloride tablet has high impurity content and has great influence on impurity detection and influence factor investigation tests of the benidipine hydrochloride tablet preparation. Therefore, a more rigorous method is required for detecting impurities in the benidipine hydrochloride tablets so as to prevent serious influence in the subsequent preparation process.

Disclosure of Invention

The invention aims to overcome the defects of the prior art, and provides a method for detecting impurities in benidipine hydrochloride tablets, which is simple to operate, reliable in method, good in repeatability and high in accuracy and is verified by methodology according to the self-component characteristics of the benidipine hydrochloride tablets.

The technical scheme adopted by the invention for solving the technical problem is as follows:

a method for detecting impurities in benidipine hydrochloride tablets adopts liquid chromatography for detection, and comprises the following steps:

(1) preparing a test solution: grinding benidipine hydrochloride tablets to powder, precisely weighing an appropriate amount of fine powder (about 25mg equivalent to benidipine hydrochloride), putting the fine powder into a 50ml measuring flask, adding an appropriate amount of methanol-diluted phosphoric acid (1 → 500) (1: 1) mixed solution, shaking up, diluting the mixed solution to a scale, shaking up, filtering, and taking a subsequent filtrate as a sample solution;

(2) preparing a reference substance solution: taking a proper amount of benidipine hydrochloride reference substance, precisely weighing, dissolving by using the mixed solution in the step (1), and quantitatively diluting to prepare a solution containing 5 mu g of benidipine hydrochloride in each 1ml as a reference substance solution;

(3) preparation of system applicability solution: taking a proper amount of benidipine hydrochloride reference substances and isomer reference substances, precisely weighing, dissolving by using the mixed solution, and quantitatively diluting to prepare a solution containing 0.5mg of benidipine hydrochloride and 5 mu g of isomer per 1ml, wherein the solution is used as a system applicability solution;

(4) detection liquid chromatography method and conditions:

respectively measuring the system applicability solution prepared in the step (3), the reference substance solution prepared in the step (2) and the test substance solution prepared in the step (1) by using a high performance liquid chromatography;

liquid chromatography conditions: chromatographic column using octadecylsilane chemically bonded silica as filler; using 0.05mol/L potassium dihydrogen phosphate solution (pH is adjusted to 3.0 by phosphoric acid) -methanol-tetrahydrofuran as mobile phase; detecting wavelength at 237nm, column temperature at 25 deg.C, adjusting flow rate to make peak retention time of benidipine hydrochloride about 20min, collecting system applicability solution 10 μ l, injecting into liquid chromatograph, and recording chromatogram until retention time of main component is 2 times;

the separation degree of the benidipine hydrochloride peak and the isomer peak is required to meet the requirement, and the separation degree is calculated by the peak area according to an external standard method;

(5) detection indexes are as follows: if an impurity peak exists in the chromatogram of the test solution, the peak area of the dehydrogenation derivative peak with the relative retention time of about 0.75 (corrected by multiplying by 1.6) is not more than 0.5 times (0.5%) of the main peak area of the control solution except the solvent peak, the peak area of other single impurities is not more than 0.2 times (0.2%) of the main peak area of the control solution, and the sum of the peak areas of the impurities is not more than 1.0% of the main peak area of the control solution.

Preferably, the column is an Agilent Pursuit, 4.6X 100mm, 3 μm or equivalent performance column.

Preferably, in the step (1), the concentration of the dilute phosphoric acid is 0.2%, and the volume ratio of the methanol to the dilute phosphoric acid is 1: 1.

Preferably, in the step (1), the shaking method is mechanical shaking for 15 min.

Preferably, in step (4), the mobile phase is a 0.05mol/L solution of potassium dihydrogen phosphate (pH adjusted to 3.0 with phosphoric acid) -methanol-tetrahydrofuran volume ratio of 60:20: 20.

The invention has the beneficial effects that:

compared with the prior art, the method for detecting the impurities in the benidipine hydrochloride tablets is simple to operate, reliable, high in repeatability and high in accuracy through methodology verification.

Detailed Description

The present invention will be further described with reference to specific examples to assist understanding of the invention. The method used in the invention is a conventional production method if no special provisions are made; the starting materials used, unless otherwise specified, are conventional commercial products.

Examples

A method for detecting impurities in benidipine hydrochloride tablets is characterized by adopting liquid chromatography for detection and comprising the following steps:

(1) preparing a test solution: grinding benidipine hydrochloride tablets to powder, precisely weighing an appropriate amount of fine powder (about 25mg equivalent to benidipine hydrochloride), placing the fine powder into a 50ml measuring flask, adding an appropriate amount of methanol-dilute phosphoric acid mixed solution (the concentration of dilute phosphoric acid is 0.2%, and the volume ratio of methanol to dilute phosphoric acid is 1: 1), mechanically shaking for 15min, shaking up, diluting the mixed solution to scale, shaking up, filtering, and taking a subsequent filtrate as a sample solution;

(2) preparing a reference substance solution: taking a proper amount of benidipine hydrochloride reference substance, precisely weighing, dissolving by using the mixed solution in the step (1), and quantitatively diluting to prepare a solution containing 5 mu g of benidipine hydrochloride in each 1ml as a reference substance solution;

(3) preparation of system applicability solution: taking a proper amount of benidipine hydrochloride reference substances and isomer reference substances, precisely weighing, dissolving by using the mixed solution, and quantitatively diluting to prepare a solution containing 0.5mg of benidipine hydrochloride and 5 mu g of isomer per 1ml, wherein the solution is used as a system applicability solution;

(4) detection liquid chromatography method and conditions:

respectively measuring the system applicability solution prepared in the step (3), the reference substance solution prepared in the step (2) and the test substance solution prepared in the step (1) by using a high performance liquid chromatography;

liquid chromatography conditions: a chromatographic column using octadecylsilane chemically bonded silica as a filler, wherein the chromatographic column is Agilent Pursuit, 4.6 multiplied by 100mm, 3 mu m or a chromatographic column with equivalent efficiency;

taking 0.05mol/L potassium dihydrogen phosphate solution (pH is adjusted to 3.0 by phosphoric acid) -methanol-tetrahydrofuran with volume ratio of 60:20:20 as a mobile phase; detecting wavelength at 237nm, column temperature at 25 deg.C, adjusting flow rate to make peak retention time of benidipine hydrochloride about 20min, collecting system applicability solution 10 μ l, injecting into liquid chromatograph, and recording chromatogram until retention time of main component is 2 times;

the separation degree of the benidipine hydrochloride peak and the isomer peak is required to meet the requirement, and the separation degree is calculated by the peak area according to an external standard method;

(5) detection indexes are as follows: if an impurity peak exists in the chromatogram of the test solution, the peak area of the dehydrogenation derivative peak with the relative retention time of about 0.75 (corrected by multiplying by 1.6) is not more than 0.5 times (0.5%) of the main peak area of the control solution except the solvent peak, the peak area of other single impurities is not more than 0.2 times (0.2%) of the main peak area of the control solution, and the sum of the peak areas of the impurities is not more than 1.0% of the main peak area of the control solution.

The impurity detection method in the benidipine hydrochloride tablets is proved to be established by methodology, and the benidipine hydrochloride tablets are good in repeatability and high in accuracy.

In conclusion, compared with the prior art, the method for detecting the impurities in the benidipine hydrochloride tablets is simple to operate and reliable, and has good repeatability and high accuracy through methodology verification.

However, the above description is only an embodiment of the present invention, and the scope of the present invention should not be limited by this, and all equivalent changes and modifications made in the claims of the present invention should be covered by the present invention.