Method for detecting dimer impurities in metoclopramide hydrochloride injection

Abstract

The invention provides a method for detecting dimer impurities in metoclopramide hydrochloride injection, which adopts high performance liquid chromatography and comprises the following steps: preparing a test solution and a system applicable solution; a chromatographic column taking octadecylsilane chemically bonded silica as a filling agent is matched with a trapping column; a mobile phase is composed of a mobile phase A and a mobile phase B, the mobile phase A is a 0.1% phosphoric acid aqueous solution, and the mobile phase B is acetonitrile; performing gradient elution; the detection wavelength is 275nm, the flow velocity is 0.9 ml / min-1. 1ml / min, the column temperature is 25-35 DEG C, and the sample injection volume is 5l; and respectively injecting the test solution and the system applicability solution into a liquid chromatograph, recording chromatograms, and calculating the content of each component. The detection method is convenient to operate and accurate in quantification, high-sensitivity and high-stability detection of the two dimer impurities can be achieved, a standard means is provided for medicine quality control, and the medicine quality control level is improved.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical analysis and detection technology, and in particular to a method for detecting dimer impurities in metoclopramide hydrochloride injection. Background Technology

[0002] Metoclopramide hydrochloride injection is a widely used gastrointestinal motility-promoting antiemetic drug in clinical practice. Its main active ingredient is metoclopramide, and the drug is a colorless, clear liquid. This drug exerts its central antiemetic effect by antagonizing dopamine D2 receptors and activating 5-HT4 receptors, while also exhibiting mild inhibition of 5-HT3 receptors. It has diverse applications in medical settings, effectively addressing vomiting symptoms induced by chemotherapy, radiotherapy, surgery, traumatic brain injury and its sequelae, and work at sea or in the air. It can also be used for symptomatic treatment of nausea and vomiting caused by acute gastroenteritis, uremia, and other conditions. Furthermore, it can assist in diagnostic duodenal intubation and barium contrast examinations, providing important support for clinical diagnosis and treatment.

[0003] The content of impurities in a drug directly affects its safety and efficacy. Metoclopramide hydrochloride injection is prone to photodegradation during storage and use, with dimer impurities being the main product of this process. Controlling the content of this impurity is a core aspect of the related substances management of metoclopramide hydrochloride injection, directly impacting the assurance of drug quality.

[0004] However, the current field of pharmaceutical quality control faces a critical technological gap: neither the Chinese Pharmacopoeia nor mainstream international pharmacopoeias include specific detection methods for this dimer impurity in the raw material and formulation standards for metoclopramide hydrochloride injection. Existing technologies for detecting metoclopramide hydrochloride injection primarily focus on determining the content of the main component or screening for other common, known impurities. The general detection methods employed cannot achieve accurate separation and quantitative analysis of the dimer impurity, failing to meet the stringent quality control requirements for impurities during drug production, storage, and clinical application. This could lead to potentially risky drugs entering the market, posing a threat to patient safety.

[0005] Therefore, developing a convenient and quantitatively accurate method for detecting dimer impurities in metoclopramide hydrochloride injection, filling the gaps in existing standards, and strengthening the quality control system of this drug has become an urgent technical problem to be solved in the field of drug analysis. Summary of the Invention

[0006] The technical problem to be solved by the present invention is to provide a convenient and quantitatively accurate method for detecting dimer impurities in metoclopramide hydrochloride injection. This method can achieve high sensitivity and high stability in detecting two kinds of dimer impurities, providing a standard technical means for impurity quality control in drug production and storage, and improving the quality control level of the injection.

[0007] The present invention solves the above-mentioned technical problems by adopting the following technical solutions: A method for detecting dimer impurities in metoclopramide hydrochloride injection, employing high-performance liquid chromatography (HPLC), comprises the following steps: (1) Solution preparation Test solution: Take metoclopramide hydrochloride injection, dilute with solvent to obtain test solution; System suitability solution: Take the dimer impurity and metoclopramide hydrochloride injection, dissolve and dilute with solvent to obtain the system suitability solution; (2) Chromatographic conditions setting The chromatographic column was packed with octadecylsilane-bonded silica gel and equipped with a trapping column; the mobile phase consisted of mobile phase A, which was a 0.1% aqueous solution of phosphoric acid, and mobile phase B, which was acetonitrile; gradient elution was used; the detection wavelength was 275 nm, the flow rate was 0.9 ml / min to 1.1 ml / min, the column temperature was 25 ℃ to 35 ℃, and the injection volume was 5 µl. (3) Sample determination Take the test solution and system suitability solution, inject them into the liquid chromatograph, record the chromatograms, and calculate the content of each component.

[0008] As one of the preferred embodiments of the present invention, in step (1), the solvent is water.

[0009] As one of the preferred embodiments of the present invention, the final concentration of metoclopramide hydrochloride in the test solution of step (1) is 1 mg / ml.

[0010] As one of the preferred embodiments of the present invention, the system suitability solution in step (1) contains two types of dimer impurities: dimer impurity 1 and dimer impurity 2. The chemical structural formulas of dimer impurity 1 and dimer impurity 2 are shown in Formulas I and II, respectively:

[0011] Formula I;

[0012] Formula II.

[0013] As one of the preferred embodiments of the present invention, in the system suitability solution, the final concentrations of dimer impurity 1 and dimer impurity 2 are 1 μg / ml, and the final concentration of metoclopramide hydrochloride is 1 mg / ml.

[0014] As one of the preferred embodiments of the present invention, in step (2), the chromatographic column is selected as Agilent XDB-C18 4.6×150mm 3.5µm, Waters XBridge-C18 4.6×150mm 3.5µm, Agilent HC-C18 4.6×150mm 5µm, or other chromatographic columns with equivalent performance.

[0015] As one of the preferred embodiments of the present invention, in step (2), the trapping column is selected as a Welch trapping column with a specification of 4.0×50mm.

[0016] As one of the preferred embodiments of the present invention, in step (2), the gradient elution program is as follows: at 0 minutes, mobile phase A accounts for 95% and mobile phase B accounts for 5%; at 14-16 minutes, mobile phase A accounts for 5% and mobile phase B accounts for 95%; at 16.1-20 minutes, mobile phase A accounts for 95% and mobile phase B accounts for 5%.

[0017] As one of the preferred embodiments of the present invention, in step (3), the chromatogram of the system suitability solution shows that the peak elution order is dimer impurity (dimer impurity 1, dimer impurity 2) and metoclopramide peak, and the resolution between each chromatographic peak is not less than 1.5.

[0018] As one of the preferred embodiments of the present invention, in step (3), the content of each component in the test sample is calculated by the area of ​​the impurity peak in the chromatogram of the test sample solution using the external standard method.

[0019] The advantages of this invention compared to the prior art are: (1) Filling a technological gap for the first time: This invention establishes a quantitative detection method for the two dimer impurities for the first time, filling the technological gap in existing standards and providing key technical support for the control of drug-related substances.

[0020] (2) Excellent separation effect and accurate quantification: This invention achieves efficient separation of dimer impurity 1 and dimer impurity 2 from the main drug component and other potential impurities by optimizing the design of exclusive chromatographic conditions (specific C18 column + trap column combination, 0.1% phosphoric acid aqueous solution-acetonitrile gradient elution system, 275nm exclusive detection wavelength, etc.). In the system suitability test, the resolution of each chromatographic peak is greater than 1.5, which effectively avoids the mutual interference between impurities and the main component and between impurities, ensuring accurate and reliable detection results and fully demonstrating the core advantage of accurate quantification.

[0021] (3) High detection sensitivity, meeting the requirements of low limit control: The detection limit of dimer impurity 1 of the present invention is as low as 0.011 μg / ml, and the quantitation limit is 0.037 μg / ml; the detection limit of dimer impurity 2 is 0.015 μg / ml, and the quantitation limit is 0.049 μg / ml; based on this, it can accurately capture trace amounts of dimer impurities in drugs, meet the quality requirements of "low limit control" of drug impurities, further ensure quantitative accuracy, and reduce the risk of drug safety.

[0022] (4) Linear stability and reliable quantitative accuracy: In the detection of the two dimer impurities in this invention, the linear relationship between the two impurities within their respective concentration ranges (impurity 1: 0.0368 μg / ml~2.228 μg / ml, impurity 2: 0.0491 μg / ml~1.986 μg / ml) is excellent, and the correlation coefficient is high. r reached 0.9992 and 0.9977 respectively, and the system has strong stability (6 consecutive injections, peak area RSD≤0.23%, retention time RSD≤0.03%), ensuring that the impurity content in different concentration ranges can be accurately quantified, avoiding the problem of large quantitative deviation in conventional methods.

[0023] (5) Easy to operate and suitable for industrial quality control: The present invention uses water as a solvent for detection, which does not require complicated pretreatment steps; the chromatographic conditions are clear and easy to repeat; the reagents used (acetonitrile, phosphoric acid) are commonly used reagents in the field of drug analysis; the chromatographic column is a commercially available general model, which does not require special instruments or consumables and is suitable for large-scale industrial quality control needs. Attached Figure Description

[0024] Figure 1 This is the chromatogram of the system suitability solution in Example 1; Figure 2 This is the chromatogram of the test solution in Example 1; Figure 3 This is a linear relationship graph of dimer impurity 1 in the experimental example; Figure 4 This is a linear relationship graph of dimer impurity 2 in the experimental example. Detailed Implementation

[0025] The embodiments of the present invention are described in detail below. These embodiments are implemented based on the technical solution of the present invention, and provide detailed implementation methods and specific operating procedures. However, the scope of protection of the present invention is not limited to the following embodiments. Unless otherwise specified, the raw materials, reagents, equipment, and methods used in the following embodiments and experimental examples are all conventional raw materials, reagents, equipment, and methods in the art, and will not be described again.

[0026] Meanwhile, the structural information of the metoclopramide hydrochloride and the dimer impurities of the present invention is shown in Table 1.

[0027] Table 1. Structural information of metoclopramide hydrochloride and various dimer impurities

[0028] Example 1 This embodiment describes a method for detecting dimer impurities in metoclopramide hydrochloride injection, employing high-performance liquid chromatography (HPLC). The steps are as follows: (1) Solution preparation Test solution: Take metoclopramide hydrochloride injection and dilute it with water to prepare a solution containing 1 mg of metoclopramide hydrochloride per 1 ml (the final concentration of metoclopramide hydrochloride is 1 mg / ml).

[0029] System suitability solution: Take dimer impurity 1, dimer impurity 2 and metoclopramide hydrochloride injection, dissolve and dilute with water to prepare a mixed solution containing 1 μg of each impurity and 1 mg of metoclopramide hydrochloride per 1 ml (the final concentrations of dimer impurity 1 and dimer impurity 2 are 1 μg / ml, and the final concentration of metoclopramide hydrochloride is 1 mg / ml).

[0030] (2) Chromatographic conditions setting An octadecylsilane-bonded silica gel column (Agilent ZORBAX Eclipse XDB-C18 column; 4.6 × 150 mm, 3.5 μm) was used as the packing material, paired with a Welch trap column (4.0 × 50 mm). The mobile phase consisted of mobile phase A (0.1% phosphoric acid aqueous solution) and mobile phase B (acetonitrile). The gradient elution program was as follows: at 0 minutes, mobile phase A accounted for 95% and mobile phase B for 5%; from 14 to 16 minutes, mobile phase A accounted for 5% and mobile phase B for 95%; from 16.1 to 20 minutes, mobile phase A accounted for 95% and mobile phase B for 5%. The detection wavelength was 275 nm, the flow rate was 1.0 ml / min, the column temperature was 30 °C, and the injection volume was 5 µl.

[0031] (3) Sample determination Take the test solution and system suitability solution, inject them into the liquid chromatograph, record the chromatograms, and calculate the content of each component in the test sample based on the impurity peak area in the chromatogram of the test solution using the external standard method.

[0032] Figure 1 Chromatogram of the solution for system suitability. Figure 2 This is the chromatogram of the test sample solution.

[0033] Depend on Figure 1 The results show that the peak order is dimer impurity 1, dimer impurity 2, and metoclopramide peak, and the resolution between dimer impurities and other impurity peaks, as well as the metoclopramide peak, is greater than 1.5, indicating that the method has good system applicability.

[0034] Depend on Figure 2 The results showed that no dimer impurities were detected in the test solution, and the sample met the requirements.

[0035] Example 2 The method for detecting dimer impurities in metoclopramide hydrochloride injection in this embodiment is basically the same as that in Example 1, except that the chromatographic column selected is a Waters XBridge-C18 (4.6×150mm 3.5µm).

[0036] Example 3 The detection method for dimer impurities in metoclopramide hydrochloride injection in this embodiment is basically the same as that in Example 1, except that the chromatographic column selected is an Agilent HC-C18 (4.6×150mm 5µm).

[0037] Example 4 The method for detecting dimer impurities in metoclopramide hydrochloride injection in this embodiment is basically the same as that in Example 1, except that the flow rate is 0.9 ml / min in the chromatographic conditions.

[0038] Example 5 The method for detecting dimer impurities in metoclopramide hydrochloride injection in this embodiment is basically the same as that in Example 1, except that the flow rate is 1.1 ml / min in the chromatographic conditions.

[0039] Example 6 The method for detecting dimer impurities in metoclopramide hydrochloride injection in this embodiment is basically the same as that in Example 1, except that the column temperature is set to 25°C in the chromatographic conditions.

[0040] Example 7 The method for detecting dimer impurities in metoclopramide hydrochloride injection in this embodiment is basically the same as that in Example 1, except that the column temperature is set to 35°C in the chromatographic conditions.

[0041] Experimental Example This experimental example provides a methodological verification of the above-mentioned method.

[0042] The instruments and chromatographic conditions are the same as in Example 1.

[0043] I. Exclusivity To ensure the specific response of the detection method to dimer impurities and to avoid interference from the main component and other potential impurities, dimer impurity 1 and dimer impurity 2 were scanned at wavelengths from 200 nm to 400 nm.

[0044] The results showed that both dimer impurities exhibited maximum absorption peaks around 275 nm. Therefore, 275 nm was selected as the specific detection wavelength for dimer impurities in this product to ensure the specificity of the detection method.

[0045] II. Limit of Detection and Limit of Quantification Accurately weigh dimer impurity 1 and dimer impurity 2, gradually dilute with water, and then accurately inject 5 µl into the liquid chromatograph and record the chromatogram. The concentration with an S / N of 10 is the limit of quantitation for the substance; the concentration with an S / N of 3 is the limit of detection for the substance. The results are shown in Tables 2 and 3.

[0046] Table 2. Results of Limit of Quantitation for Dimer Impurities

[0047] Table 3. Results of detection limits for dimer impurities

[0048] The results above show that the method of the present invention has high sensitivity.

[0049] III. Linearity and Range Accurately weigh dimer impurity 1 and dimer impurity 2, dissolve them in water, and dilute them to prepare linear solutions with concentrations of 0.25 μg / ml, 0.50 μg / ml, 1.00 μg / ml, 1.50 μg / ml, and 2.00 μg / ml, respectively. Under the chromatographic conditions of Example 1, accurately measure 5 μl of each solution and inject it into the liquid chromatograph, recording the chromatograms. Simultaneously, use the limit of quantitation concentration as the linear starting point, and plot a standard curve with peak area on the ordinate and concentration on the abscissa. The results are as follows: Figure 3 , 4 As shown in Tables 4 and 5.

[0050] Table 4. Experimental results of linear relationship of dimeric impurity 1

[0051] Table 5. Experimental results of linear relationship between dimeric impurity 2

[0052] The results above show that dimer impurity 1 exhibits good linearity in the concentration range of 0.0368 µg / ml to 2.228 µg / ml, and dimer impurity 2 exhibits good linearity in the concentration range of 0.0491 µg / ml to 1.986 µg / ml.

[0053] IV. System Applicability Dimer impurity 1, dimer impurity 2, and metoclopramide hydrochloride injection were dissolved and diluted with water to prepare a mixed solution containing 1.0 mg of metoclopramide hydrochloride and 1 μg of each impurity per 1 ml. This solution was used as the system suitability solution. Six consecutive injections were performed, and the chromatograms were recorded to examine the system suitability of this method.

[0054] The results are shown in Table 6.

[0055] Table 6. Results of System Suitability Test

[0056] The results above show that the separation between each impurity peak and before and after the main peak is greater than 1.5, the RSD of each peak area is less than 2.0%, and the RSD of the retention time is less than 1.0%, indicating that the system has good applicability.

[0057] V. Solution stability 1. Stability of the test solution Accurately measure metoclopramide hydrochloride injection solution and dilute with water to prepare a solution containing 1.0 mg of metoclopramide hydrochloride per 1 ml, which is used as the test solution. The solution is left at room temperature and injected at 0 h, 2 h, 4 h, 6 h, 8 h, 10 h, 12 h, 18 h, and 24 h to investigate its stability. The results are shown in Table 7.

[0058] Table 7. Results of the stability test of the test sample solution

[0059] The results above show that the test solution of the present invention has good stability within 24 hours.

[0060] 2. Stability of the mixed impurity control solution Accurately weigh dimer impurity 1 and dimer impurity 2, dissolve and dilute them in water to prepare a solution containing approximately 1 µg of each impurity per 1 ml. Incubate at room temperature and inject at 0 h, 2 h, 4 h, 6 h, 8 h, 10 h, 12 h, 18 h, and 24 h to examine the stability of the mixed impurity control solution. The results are shown in Table 8.

[0061] Table 8. Results of stability test on the mixed impurity control solution

[0062] The results above show that the mixed impurity control solution has good stability within 24 hours.

[0063] VI. Accuracy Accurately weigh dimer impurity 1 and dimer impurity 2, dissolve and dilute them in water to prepare a solution containing 1 µg of each impurity per 1 ml, which is used as a mixed impurity reference solution.

[0064] According to the impurity addition amounts of 50% (0.5µg / ml), 100% (1µg / ml) and 150% (1.5µg / ml) standards, respectively, appropriate amounts of each mixed impurity reference solution were added to metoclopramide hydrochloride injection, 3 portions of each concentration, and shaken well to obtain the test solution.

[0065] Under the chromatographic conditions and detection methods described above, the content of each impurity in each test sample was calculated based on peak area using the external standard method, and the recovery rate was calculated. The results are shown in Tables 9 and 10.

[0066] Table 9. Results of the recovery test for dimeric impurity 1

[0067] Table 10. Results of the recovery test for dimeric impurity 2

[0068] The results above show that the average recovery rate of each impurity determined by the method of the present invention is between 80% and 120%, indicating that the method has high accuracy in determining each impurity.

[0069] VII. Durability The system suitability solution was used as the robustness solution. The tolerance of this chromatographic condition was investigated by changing the column temperature, flow rate, and chromatographic columns from different manufacturers within a certain range. The results are shown in Tables 11-13.

[0070] Table 11. Durability test results (at different column temperatures)

[0071] Table 12. Durability test results (at different flow rates)

[0072] Table 13. Durability test results (different chromatographic columns)

[0073] The results above show that the resolution meets the requirements when the column temperature is varied from 25℃ to 35℃, the flow rate is varied from 0.9 ml / min to 1.1 ml / min, and the columns are from different manufacturers. This method has good robustness.

[0074] In summary, the detection method provided by this invention has high specificity and good durability. Changing the chromatographic conditions within a small range will not affect the detection of dimer impurities. The method provided by this invention can accurately detect dimer impurities in metoclopramide hydrochloride injection, with good separation between impurities and between the peaks before and after the main component. The detection results are accurate and reliable, and can more scientifically control the quality of metoclopramide hydrochloride injection.

[0075] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for detecting dimer impurities in a meclizine hydrochloride injection, comprising: The high performance liquid chromatography method is as follows: (1) Solution preparation Test solution: Take methoxychlorpromazine hydrochloride injection, dilute with solvent to obtain test solution; System suitability solution: Take dimer impurities and methoxychlorpromazine hydrochloride injection, dissolve and dilute with solvent to obtain system suitability solution; (2) Chromatographic condition setting The chromatographic column with octadecylsilane bonded silica gel as filler is matched with a trapping column; the mobile phase is composed of mobile phase A and mobile phase B, mobile phase A is 0.1% phosphoric acid aqueous solution, and mobile phase B is acetonitrile; gradient elution; detection wavelength is 275 nm, flow rate is 0.9 ml / min~1.1 ml / min, column temperature is 25℃~35℃, and injection volume is 5µl; (3) Sample determination Take the test solution and the system suitability solution, respectively, and inject them into the liquid chromatograph, record the chromatogram, and calculate the content of each component.

2. The method for detecting dimer impurities in methoclopramide hydrochloride injection according to claim 1, characterized by, In the step (1), the solvent is water.

3. The method for detecting dimer impurities in methoclopramide hydrochloride injection according to claim 1, characterized by, In the test solution of the step (1), the final concentration of methoxychlorpromazine hydrochloride is 1 mg / ml.

4. The method for detecting dimer impurities in a meclizine hydrochloride injection according to claim 1, wherein In the system suitability solution of the step (1), the dimer impurities include dimer impurity 1 and dimer impurity 2; the chemical structural formulas of the dimer impurity 1 and the dimer impurity 2 are shown in the following formula I and II, respectively: Formula I; Formula II.

5. The method for detecting dimer impurities in a meclizine hydrochloride injection according to claim 4, wherein In the system suitability solution, the final concentrations of the dimer impurity 1 and the dimer impurity 2 are 1µg / ml respectively, and the final concentration of methoxychlorpromazine hydrochloride is 1mg / ml.

6. The method for detecting dimer impurities in a meclizine hydrochloride injection according to claim 1, wherein, In the step (2), the chromatographic column is selected from Agilent XDB-C18 4.6×150mm 3.5µm, Waters XBridge-C18 4.6×150mm 3.5µm or Agilent HC-C18 4.6×150mm 5µm.

7. The method for detecting dimer impurities in a meclizine hydrochloride injection according to claim 1, wherein In the step (2), the trapping column is selected from Welch trapping column with a specification of 4.0×50mm.

8. The method for detecting dimer impurities in a meclizine hydrochloride injection according to claim 1, wherein, In the step (2), the gradient elution program is as follows: at 0 minutes, the proportion of mobile phase A is 95% and the proportion of mobile phase B is 5%; at 14~16 minutes, the proportion of mobile phase A is 5% and the proportion of mobile phase B is 95%; at 16.1~20 minutes, the proportion of mobile phase A is 95% and the proportion of mobile phase B is 5%.

9. The method for detecting dimer impurities in a meclizine hydrochloride injection according to claim 1, wherein, In the step (3), in the chromatogram of the system suitability solution, the peak order is dimer impurity, methoxychlorpromazine peak in turn, and the separation degree between each chromatographic peak is not less than 1.

5.

10. The method for detecting dimer impurities in a meclizine hydrochloride injection according to claim 1, wherein In the step (3), the content of each component in the test solution is calculated according to the impurity peak area in the chromatogram of the test solution by external standard method.

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