A method for detecting sulfonic acid epinephrine impurities in epinephrine drugs

Abstract

The application provides a detection method for a sulfonic acid adrenaline impurity in an adrenaline drug, in particular to a method for detecting a sulfonic acid adrenaline impurity in an adrenaline injection solution by HPLC-ECD, compared with the prior art, the technical scheme of the application has significant advantages in sensitivity, stability, accuracy and the like, and the detection method provided by the application scheme is suitable for adrenaline single agent and / or compound including adrenaline and the like, and has important significance for improving the product quality of the adrenaline drug.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical analysis, and specifically to a method for detecting sulfonate epinephrine impurities in epinephrine-type drugs, specifically a method for detecting sulfonate epinephrine impurities in epinephrine-type injections by HPLC-ECD. Background Technology

[0002] Adrenaline is a catecholamine neurotransmitter that is easily deactivated by oxidation, demethylation, or reaction with excipients in formulations. Adrenaline readily reacts with sulfite ions generated by the dissociation of sodium metabisulfite in water to form adrenaline sulfonate (reaction shown in the figure below).

[0003]

[0004] Existing detection methods for the aforementioned impurities have certain shortcomings. For example, the Chinese Pharmacopoeia does not control the content of epinephrine sulfonate, and while some drug import registration standards provide detection methods for epinephrine sulfonate content, these methods have limited sensitivity and fail to meet separation requirements, leaving room for optimization. Chinese Patent 202211675677.6 discloses a detection method for epinephrine sulfonate, but analysis of the results obtained from reproducing this method revealed accuracy defects. Chinese Patent 202311361555.4 discloses a detection method for impurities in epinephrine injection solution, but analysis of the results obtained from reproducing this method revealed insufficient sensitivity.

[0005] When the content of epinephrine sulfonate impurities in adrenaline drugs is too high, it will affect the therapeutic effect of the active ingredient. The existing detection methods are insufficient to meet the actual needs. Therefore, it is necessary to develop a new detection method to deal with the detection of epinephrine sulfonate impurities in adrenaline drugs (adrenaline monotherapy, and / or combination drugs including epinephrine, etc.). Summary of the Invention

[0006] The purpose of this invention is to overcome the shortcomings of the prior art and provide a method for detecting epinephrine sulfonate that is widely applicable, safe in the measurement process, low in cost, and accurate and reliable in the measurement results.

[0007] The specific details of the invention are as follows.

[0008] A method for detecting epinephrine sulfonate impurities in adrenaline-based drugs, the method being applicable to drugs containing epinephrine alone and / or compound drugs including epinephrine.

[0009] In some embodiments, a method for detecting epinephrine sulfonate impurities in epinephrine-based drugs is provided, the method being applicable to epinephrine injection, isoproterenol injection, norepinephrine bitartrate injection, lidocaine epinephrine injection, articaine epinephrine injection, and other compound preparations containing epinephrine.

[0010] A method for detecting epinephrine sulfonate impurities in adrenaline-based drugs, the method comprising: preparing a solution, setting chromatographic conditions, and preparing a mobile phase.

[0011] In some embodiments, a method for detecting epinephrine sulfonate impurities in adrenaline-based drugs includes a solution preparation step in which the solvent is a metabisulfite solution, such as sodium metabisulfite solution or potassium metabisulfite solution.

[0012] In some embodiments, a method for detecting epinephrine sulfonate impurities in adrenaline-based drugs includes a solution preparation step in which a potassium metabisulfite solution is preferably used as the solvent.

[0013] In some implementations, a method for detecting epinephrine sulfonate impurities in adrenaline-based drugs includes steps for setting chromatographic conditions such as column selection, column temperature adjustment, injection volume selection, and detector settings.

[0014] In some embodiments, a method for detecting sulfonate adrenaline impurities in adrenaline-based drugs is provided, wherein the chromatographic column used in the method may be a C8 column or a C18 column, with a C18 column being preferred.

[0015] In some embodiments, a method for detecting sulfonate epinephrine impurities in adrenaline-based drugs is provided, wherein the chromatographic column used in the method is a C18 column, wherein the specifications of the C18 column can be 4.6 mm × 250 mm, 5 μm; 4.0 mm × 250 mm, 3 μm; or 4.0 mm × 250 mm, 10 μm.

[0016] In some embodiments, a method for detecting epinephrine sulfonate impurities in adrenaline-based drugs is provided, wherein the chromatographic column used in the method is a C18 column, wherein the C18 column can be used alone or two columns can be used in series.

[0017] In some embodiments, a method for detecting epinephrine sulfonate impurities in adrenaline-based drugs is provided, wherein the chromatographic column is used in a series of two columns, which may be two C8 columns or two C18 columns connected in series.

[0018] In some embodiments, a method for detecting epinephrine sulfonate impurities in adrenaline-based drugs is provided, wherein the chromatographic column is used in a series of two columns, preferably two C18 columns.

[0019] In some embodiments, a method for detecting epinephrine sulfonate impurities in adrenaline-based drugs includes a column temperature range of 25-40°C, preferably 30°C, in the step of setting chromatographic conditions.

[0020] In some embodiments, a method for detecting epinephrine sulfonate impurities in adrenaline-based drugs includes an injection volume of 25 μl in the step of setting chromatographic conditions.

[0021] In some embodiments, a method for detecting epinephrine sulfonate impurities in adrenaline-based drugs may include an electrochemical detector or a fluorescence detector in the step of setting chromatographic conditions.

[0022] In some embodiments, a method for detecting epinephrine sulfonate impurities in adrenaline-based drugs, wherein the detector in the step of setting chromatographic conditions is preferably an electrochemical detector.

[0023] In some embodiments, a method for detecting epinephrine sulfonate impurities in adrenaline-based drugs, wherein the detector in the step of setting chromatographic conditions is preferably an amperometric detector from the category of electrochemical detectors.

[0024] In some embodiments, a method for detecting epinephrine sulfonate impurities in adrenaline-based drugs, wherein the detector parameters in the step of setting chromatographic conditions are set as follows: Detector: Ampere detector; Detector temperature: 30℃; Detection electrode: glassy carbon electrode; Reference electrode: Silver-silver chloride electrode; Electrode voltage: +650mV.

[0025] In some embodiments, a method for detecting sulfonate epinephrine impurities in adrenaline-based drugs includes a mobile phase preparation process as follows: glacial acetic acid is taken, water is added and mixed, the pH value is adjusted, and sodium heptanesulfonate, disodium ethylenediaminetetraacetate solution, and potassium chloride are added and mixed.

[0026] In some embodiments, a method for detecting sulfonate epinephrine impurities in adrenaline-based drugs includes a mobile phase preparation process as follows: glacial acetic acid is taken, water is added and mixed, the pH value is adjusted, sodium octane sulfonate, disodium ethylenediaminetetraacetate solution, and potassium chloride are added and mixed.

[0027] In some embodiments, a method for detecting epinephrine sulfonate impurities in adrenaline-based drugs is provided, wherein the mobile phase pH ranges from 3 to 4.

[0028] In some embodiments, a method for detecting epinephrine sulfonate impurities in adrenaline-based drugs is provided, wherein the pH of the mobile phase in the method is in the range of 3.3-3.6.

[0029] This invention establishes for the first time a method for detecting sulfonate epinephrine impurities in epinephrine-type drugs, overcoming a series of technical difficulties in the prior art, such as low sensitivity, poor selectivity and poor repeatability.

[0030] The beneficial effects of this invention compared to the prior art are as follows.

[0031] (1) The detection method of the present invention can be applied to the detection of adrenaline sulfonate in adrenaline drugs, and is applicable to adrenaline monotherapy preparations, compound preparations including adrenaline, and other forms including adrenaline.

[0032] (2) The method of using tandem chromatographic columns in the present invention increases the theoretical plate number of the chromatographic system compared with the prior art, thereby enhancing the separation ability between sulfonate adrenaline and the main component, and improving its peak shape and response stability.

[0033] (3) In this invention, sodium heptanesulfonate is introduced into the mobile phase as an ion-pairing reagent to complement the scheme in (2), and the pH of the mobile phase is controlled in the range of 3.2 to 3.6 so that adrenaline sulfonate forms a stable ion pair in the chromatographic system, thereby improving its retention behavior in the reversed phase system and increasing the resolution, and reducing the risk of co-elution.

[0034] (4) The disodium ethylenediaminetetraacetate chelating agent used in this invention, in combination with the schemes in (2) and (3), hinders the oxidation of catechol structure, reduces baseline drift and burr generation, improves the stability of chromatographic peaks, and ensures accurate quantification of trace impurities.

[0035] (5) In this invention, an electrochemical detector is used in conjunction with the schemes in (2), (3), and (4), and the detection potential is controlled within the range of 0.60V to 0.70V to obtain the best electrochemical response of sulfonate epinephrine. While ensuring detection sensitivity, the background noise is reduced. At the same time, the DC mode is used to achieve high selectivity detection of sulfonate epinephrine and complete separation of sulfonate epinephrine from the main component, thus solving the problem of poor repeatability of sulfonate epinephrine.

[0036] (6) This invention uses the principal component external standard method for detection, resulting in more reliable results. Compared with the prior art, this invention is simple to operate, reliable, highly sensitive, and selective. Establishing an efficient detection method for adrenaline sulfonate helps in product quality control and process optimization, and is suitable for rapid quality control in industrial production, which is of great significance for improving product quality. Attached Figure Description

[0037] Appendix Figure 1 The image shows the chromatogram of epinephrine sulfonate as determined in Example 1. Detailed Implementation

[0038] To better illustrate the technical solution of the invention, the following embodiments and comparative examples will be used for further explanation. The instruments, reagents, and other materials involved in the following experiments can all be obtained through conventional commercial channels.

[0039] It should be clarified that the content described in this section is only a part of the invention, not all of it. Therefore, all other technical solutions obtained by those skilled in the art based on the technical solutions of this invention without inventive effort are within the scope of protection of this invention. Example 1 (Determination of sulfonate epinephrine in articaine epinephrine injection)

[0040] (1) Preparation of solution: Solvent: 0.05% potassium metabisulfite solution; Reference solution: Weigh approximately 2 mg of adrenaline reference standard accurately, place it in a 200 ml volumetric flask, dissolve and dilute to the mark with solvent, and shake well to prepare the reference standard stock solution; accurately measure 1 ml of the reference standard stock solution, place it in a 20 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0041] Test solution: Take this product (specification: 1.7ml: 68mg articaine hydrochloride and 17μg epinephrine tartrate (calculated as epinephrine)) to obtain the solution.

[0042] (2) Chromatographic conditions: Chromatographic column: C18 4.6mm×250mm, 5μm (two identical columns connected in series) Column temperature: 30℃ Injection volume: 25 μl Detector: Ampere detector Detector temperature: 30℃ Detection electrode: glassy carbon electrode Reference electrode: Silver-silver chloride electrode Electrode voltage: +650mV Mobile phase: Take 50 ml of glacial acetic acid, add 930 ml of water, mix well, adjust the pH value to 3.4 with 50% sodium hydroxide solution, add 1.2 g of sodium heptanesulfonate, 1 ml of 0.1 mol / L disodium ethylenediaminetetraacetate solution, and 0.35 g of potassium chloride, and mix well.

[0043] Runtime: 45 min Washing method: isocratic washing (3) Sample injection detection After the system stabilizes, inject one syringe of solvent, five syringes of reference solution, and one syringe of test solution, and record the chromatogram (see attached). Figure 1 ).

[0044] (4) Measurement results The content of epinephrine sulfonate in the epinephrine injection is less than 7.5%, and the separation degree between epinephrine and epinephrine sulfonate is 51.96, which is greater than 1.5, indicating good separation. The theoretical plate number is 30,000, indicating that the peak shape of epinephrine sulfonate is good. Example 2 Methodological Validation

[0045] Exclusivity Solvent: 0.05% potassium metabisulfite solution.

[0046] Excipient solution: Prepare an adrenaline-free solution with the same concentration as the prescription.

[0047] Reference solution: Weigh approximately 2 mg of adrenaline reference standard accurately, place it in a 200 ml volumetric flask, dissolve and dilute to the mark with solvent, and shake well to prepare the reference standard stock solution; accurately measure 1 ml of the reference standard stock solution, place it in a 20 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0048] Stock solution of adrenaline sulfonate reference standard: Weigh approximately 2 mg of adrenaline sulfonate reference standard accurately, place it in a 20 ml volumetric flask, dissolve and dilute to the mark with solvent, and shake well to prepare the stock solution of adrenaline sulfonate; Accurately measure 3.75 ml of the stock solution of adrenaline sulfonate, place it in a 50 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0049] Epinephrine sulfonate reference solution: Accurately measure 1 ml of epinephrine sulfonate reference solution into a 10 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0050] Spiked test solution: Accurately measure 1 ml of epinephrine sulfonate reference stock solution, place it in a 10 ml volumetric flask, dilute to the mark with the sample, and shake well.

[0051] The chromatographic conditions were the same as in Example 1.

[0052] The solvent, excipient solution, reference solution, adrenaline sulfonate reference solution, and spiked test solution were injected separately for testing. The specific test results are shown in the table below.

[0053] Table 1: Results of Specificity Measurement

[0054] Conclusion: The solvent does not interfere with the determination of epinephrine sulfonate. In the spiked test solution, epinephrine sulfonate achieves baseline separation from adjacent peaks. This invention demonstrates good specificity for detecting the epinephrine impurity epinephrine sulfonate. Even when other potentially interfering components are present in the sample, the analytical method can still correctly identify and determine the target analyte.

[0055] (2) Limit of quantitation and limit of detection Solvent: 0.05% potassium metabisulfite solution.

[0056] Reference stock solution: Weigh approximately 2 mg of adrenaline reference standard accurately, place it in a 200 ml volumetric flask, dissolve and dilute to the mark with solvent, and shake well to prepare the reference stock solution.

[0057] Epinephrine sulfonate stock solution: Weigh approximately 2 mg of epinephrine sulfonate reference standard accurately, place it in a 20 ml volumetric flask, dissolve and dilute to the mark with solvent, shake well, and use as epinephrine sulfonate stock solution. Limit of Quantitation (LOQ) Stock Solution: Accurately measure 7.5 ml of epinephrine sulfonate stock solution and 10 ml of epinephrine reference stock solution, place them in the same 100 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0058] Limit of Quantitation Solution: Accurately measure 0.2 ml of the limit of quantitation stock solution, place it in a 20 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0059] Detection limit solution: Accurately measure 5 ml of the quantitation limit solution, place it in a 10 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0060] The chromatographic conditions were the same as in Example 1.

[0061] The limit of quantitation (LOQ) solution and the limit of detection (LOD) solution were injected separately for determination. In the LQ solution, the signal-to-noise ratio (SNR) of epinephrine sulfonate should be no less than 10:1; in the LOD solution, the SNR of epinephrine sulfonate should be no less than 3:1; the retention time and the RSD of the response should both be less than 10%. The determination results are shown in the table below.

[0062] Table 2: Results of determination of limit of quantitation and limit of detection

[0063] Conclusion: In the limit of quantitation solution, the signal-to-noise ratio of sulfonate epinephrine is greater than 10:1. Sulfonate epinephrine concentrations of 0.153 μg / ml and above can be well quantitatively detected, with an RSD of retention time of less than 1.0% and an RSD of peak area of ​​less than 10%. The signal-to-noise ratio of sulfonate epinephrine in the limit of detection solution is greater than 3:1, indicating that the method of the present invention has high sensitivity.

[0064] (3) Linear Solvent: 0.05% potassium metabisulfite solution.

[0065] Reference stock solution: Weigh approximately 2 mg of adrenaline reference standard accurately, place it in a 200 ml volumetric flask, dissolve and dilute to the mark with solvent, and shake well to prepare the reference stock solution.

[0066] Stock solution of adrenaline sulfonate reference standard: Weigh about 2 mg of adrenaline sulfonate reference standard accurately, place it in a 20 ml volumetric flask, add solvent to dissolve and dilute to the mark, shake well, and use it as the stock solution of adrenaline sulfonate. Linear stock solution: Accurately measure 3.75 ml of epinephrine sulfonate stock solution and 5 ml of epinephrine reference stock solution, place them in the same 50 ml volumetric flask, dilute to the mark with solvent, and shake well to obtain the linear stock solution.

[0067] Linear solution 1: Accurately measure 0.2 ml of the linear stock solution, place it in a 20 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0068] Linear solution 2: Accurately measure 0.4 ml of the linear stock solution, place it in a 20 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0069] Linear solution 3: Accurately measure 1 ml of the linear stock solution, place it in a 20 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0070] Linear solution 4: Accurately measure 1 ml of the linear stock solution, place it in a 10 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0071] Linear solution 5: Accurately measure 1.5 ml of the linear stock solution, place it in a 10 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0072] Linear solution 6: Accurately measure 2 ml of the linear stock solution, place it in a 10 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0073] Linear solution 7: Accurately measure 6 ml of adrenaline reference stock solution, place it in a 50 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0074] Linear solution 8: Accurately measure 2 ml of adrenaline reference stock solution, place it in a 50 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0075] Linear solution 9: Accurately measure 2.5 ml of adrenaline reference stock solution, place it in a 50 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0076] Linear solution 10: Accurately measure 3 ml of adrenaline reference stock solution, place it in a 50 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0077] Inject each linear solution separately, record the chromatogram, and perform linear regression using the least squares method with concentration as the x-axis and peak area of ​​each linear gradient solution as the y-axis. Calculate the correlation coefficient, slope, and intercept. The linear correlation coefficient r should not be less than 0.99. The linear determination results are shown in the table below.

[0078] Table 3: Linearity Measurement Results

[0079] Conclusion: Within the linear range, the linear correlation coefficients (r) of sulfonate adrenaline and adrenaline are both greater than 0.99, indicating that the present invention has good linearity in detecting sulfonate adrenaline within adrenaline.

[0080] (4) Repeatability Solvent: 0.05% potassium metabisulfite solution.

[0081] Reference solution: Weigh approximately 2 mg of adrenaline reference standard accurately, place it in a 200 ml volumetric flask, dissolve and dilute to the mark with solvent, and shake well to prepare the reference standard stock solution; accurately measure 1 ml of the reference standard stock solution, place it in a 20 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0082] Test solution: Take this product (specification: 1.7ml: 68mg articaine hydrochloride and 17μg epinephrine tartrate (calculated as epinephrine)) to obtain the solution.

[0083] This was achieved by determining the RSD of epinephrine sulfonate in six test solutions. The requirement was that the RSD of epinephrine detection in the six spiked test solutions be less than 5%. The repeatability test results are shown in the table below: Table 4: Repeatability Test Results

[0084] Conclusion: The RSD of epinephrine sulfonate in the 6 test solutions was 2.34%, which met the requirements. The repeatability of this invention in detecting epinephrine sulfonate in epinephrine is good.

[0085] (5) Accuracy Solvent: 0.05% potassium metabisulfite solution.

[0086] Stock solution of adrenaline sulfonate reference standard: Weigh approximately 2 mg of adrenaline sulfonate reference standard accurately, place it in a 20 ml volumetric flask, dissolve and dilute to the mark with solvent, and shake well to prepare the stock solution of adrenaline sulfonate; Accurately measure 3.75 ml of the stock solution of adrenaline sulfonate, place it in a 50 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0087] Epinephrine sulfonate reference solution: Accurately measure 1 ml of epinephrine sulfonate reference solution into a 10 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0088] Test solution: Take this product (specification: 1.7ml: 68mg articaine hydrochloride and 17μg epinephrine tartrate (calculated as epinephrine)) to obtain the solution.

[0089] 10% Spiked Test Solution: Accurately measure 0.1 ml of epinephrine sulfonate reference stock solution, place it in a 10 ml volumetric flask, dilute to the mark with the sample, and shake well. Prepare 3 parallel solutions.

[0090] 100% spiked test solution: Accurately measure 1 ml of epinephrine sulfonate reference stock solution, place it in a 10 ml volumetric flask, dilute to the mark with the sample, and shake well. Prepare 3 parallel solutions.

[0091] 100% spiked test solution: Accurately measure 1.5 ml of epinephrine sulfonate reference stock solution, place it in a 10 ml volumetric flask, dilute to the mark with the sample, and shake well. Prepare 3 parallel aliquots.

[0092] The chromatographic conditions were the same as in Example 1. The sample was injected and detected, and the chromatogram was recorded. The accuracy test results are shown in the table below.

[0093] Table 5: Accuracy Measurement Results

[0094] Conclusion: The average recovery rate of epinephrine sulfonate detected by the method of the present invention was 105.6%, and the RSD was less than 10%, indicating that the detection method of the present invention has good accuracy.

[0095] (6) Durability Solvent: 0.05% potassium metabisulfite solution.

[0096] Reference solution: Weigh approximately 2 mg of adrenaline reference standard accurately, place it in a 200 ml volumetric flask, dissolve and dilute to the mark with solvent, and shake well to prepare the reference standard stock solution; accurately measure 1 ml of the reference standard stock solution, place it in a 20 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0097] Test solution: Take this product (specification: 1.7ml: 68mg articaine hydrochloride and 17μg epinephrine tartrate (calculated as epinephrine)) to obtain the solution.

[0098] The chromatographic conditions differed from those in Example 1 in that the flow rates were adjusted to 0.8 ml / min and 1.2 ml / min, the column temperatures to 25°C and 35°C, and the sodium heptanesulfonate concentrations to 95% and 105%, respectively. The changes in the detection amount of the test sample solution were examined. The robustness test results are shown in the table below: Table 6: Durability Test - Flow Rate Variation Measurement Results

[0099] Table 7: Durability Test - Column Temperature Change Measurement Results

[0100] Table 8: Durability Test - Results of Measurement of Change in Sodium Heptanesulfonate Proportion in Mobile Phase

[0101] Conclusion: Under various conditions of flow rate variation ±0.2 ml / min, column temperature variation ±5℃, and heptanesulfonate sodium variation ±5%, the detection concentration of epinephrine in the test solution changed by less than 10%, indicating good robustness. Comparative Example 1

[0102] This comparative example uses fluorescence detection.

[0103] Solvent: 0.05% sodium metabisulfite solution (dissolve 0.5g of sodium metabisulfite in 1000ml of water).

[0104] Test solution: Take 5 ml of commercially available adrenaline injection, mix well, and the solution is ready.

[0105] Reference solution: Weigh approximately 2.5 mg of epinephrine sulfonate reference standard accurately, place it in a 100 ml volumetric flask, add an appropriate amount of solvent, sonicate to dissolve, then dilute to the mark with solvent and shake well to prepare epinephrine sulfonate stock solution. Accurately measure 3 ml of epinephrine sulfonate stock solution, place it in a 100 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0106] 20% spiked test solution: Accurately measure 0.06 ml of epinephrine sulfonate stock solution and place it in a 10 ml volumetric flask. Dilute to the mark with the test solution and mix well. Prepare 3 parallel solutions.

[0107] 100% spiked test solution: Accurately measure 0.3 ml of epinephrine sulfonate stock solution and place it in a 10 ml volumetric flask. Dilute to the mark with the test solution and mix well. Prepare 3 parallel solutions.

[0108] 150% spiked test solution: Accurately measure 0.45 ml of epinephrine sulfonate stock solution and place it in a 10 ml volumetric flask. Dilute to the mark with the test solution and mix well. Prepare 3 parallel solutions.

[0109] The chromatographic conditions are as follows: Column: Welch Ultimate XB-C18 4.6mm × 250mm, 5μm The fluorescence detector has the following detection wavelengths: excitation wavelength 280nm, emission wavelength 320nm; Column temperature: 25℃ Injection volume: 20 μl Sample chamber temperature: 4℃ Flow rate: 0.8 ml / min Mobile phase A: 1.1% sodium heptanesulfonate solution (dissolve approximately 11.021 g of sodium heptanesulfonate in 1000 ml of water, and adjust the pH to 2.5 with 1 mol / L phosphoric acid solution). Mobile phase B: Acetonitrile Washing procedure: Table 9: Elution procedure for Comparative Example 1

[0110] Sample injection and detection were performed, and the spectra were recorded. The experimental results are shown in the table below: Table 10: Accuracy Measurement Results of Comparative Example 1

[0111] Conclusion: Compared with the accuracy test results in Example 1 (5), the recovery rate of sulfonate epinephrine was significantly higher when the fluorescence detector was used to detect sulfonate epinephrine, indicating that the HPLC-ECD method is more accurate and reliable. Comparative Example 2

[0112] The solution preparation and chromatographic conditions were the same as in Example 1, except that a C18 4.6mm × 250mm, 5μm single column was used. The solvent, reference solution, and test solution were injected separately for testing, and the specific results are shown in the table below.

[0113] Table 11: Results of Repeatability Tests in Comparative Example 2

[0114] Conclusion: When comparing the repeatability test results of the single column detection of sulfonate epinephrine using an electrochemical detector with those of Example 1 using two C18 columns in series (4), the detection amount of sulfonate epinephrine gradually decreased, with an RSD of 14%, indicating that the peak area response was unstable. This suggests that the repeatability of the two C18 columns in series method for detecting sulfonate epinephrine is better and the results are more reliable. (Methods for determining adrenaline content in imported pharmaceutical standards)

[0115] The solution preparation and chromatographic conditions were the same as in Example 1, except that a Merk Lichrospher 100RP C8 4.0mm × 250mm, 5μm single column was used. The solvent, reference solution, and test solution were injected separately for testing, and the specific results are shown in the table below.

[0116] Table 12: Results of Comparative Example 3 - Specificity Determination Results

[0117] Conclusion: Comparing the results of the detection of epinephrine sulfonate using a single C8 column with an electrochemical detector, the minimum resolution between epinephrine sulfonate and adjacent peaks in the test solution was 1.41, which is less than 1.5, indicating poor specificity. This suggests that the C18 column provides good specificity for the detection of epinephrine sulfonate. Comparative Example 4

[0118] The solution preparation and chromatographic conditions were the same as in Example 1, except that two C18 4.0mm×250mm, 3μm columns were used in series. The solvent, reference solution, and test solution were injected separately for testing, and the specific results are shown in the table below.

[0119] Table 13: Results of Comparative Example 4 - Repeatability Test Results

[0120] Conclusion: Comparing the results of the assay using two 3µm C18 columns in tandem with the results of Example 1, the RSD of the detected amount of epinephrine in the test solution was 13.44%, which is relatively large. This indicates that the precision of the assay using two 5µm C18 columns in tandem for the detection of epinephrine is good. Comparative Example 5

[0121] The solution preparation and chromatographic conditions were the same as in Example 1, except that two 10μm C18 4.0mm×250mm columns were used in series. The solvent, reference solution, and test solution were injected separately for testing, and the specific results are shown in the table below.

[0122] Table 14: Results of Comparative Example 5 - Repeatability Test Results

[0123] Conclusion: Comparing the results of the detection of epinephrine sulfonate using two 10µm C18 columns in series with an electrochemical detector, the RSD of the epinephrine sulfonate detection in the test solution was 10.47%, which is relatively large. This indicates that the 5µm C18 column provides good precision for the detection of epinephrine sulfonate.

[0124] This invention establishes for the first time a detection method for epinephrine sulfonate impurities in adrenaline-containing drugs. It is applicable to the detection of epinephrine sulfonate in adrenaline-containing drugs, including single-agent epinephrine preparations, compound preparations containing epinephrine, and other forms containing epinephrine. This method is of great significance for improving the product quality of adrenaline-containing drugs.

Claims

1. A method for detecting epinephrine sulfonate impurities in adrenaline-based drugs, characterized in that, The adrenaline-based drugs include single-agent preparations of adrenaline or compound preparations containing adrenaline.

2. The method for detecting epinephrine sulfonate impurities in adrenaline-based drugs according to claim 1, characterized in that, The adrenaline-like drugs are compound preparations containing adrenaline.

3. The method for detecting epinephrine sulfonate impurities in adrenaline-based drugs according to claim 1, characterized in that, The detection method includes: preparing a solution, setting chromatographic conditions, and preparing a mobile phase.

4. The method for detecting epinephrine sulfonate impurities in adrenaline-based drugs according to claim 1, characterized in that, Setting chromatographic conditions in the detection method includes: selecting a chromatographic column, setting instrument conditions, and selecting a detector.

5. The method for detecting epinephrine sulfonate impurities in adrenaline-based drugs according to claim 4, characterized in that, The chromatographic columns that can be selected in setting the chromatographic conditions are: C8 column and C18 column, with C18 column being preferred.

6. The method for detecting epinephrine sulfonate impurities in adrenaline-based drugs according to claim 4, characterized in that, The chromatographic column selected in the chromatographic conditions settings is a C18 column.

7. The method for detecting epinephrine sulfonate impurities in adrenaline-based drugs according to claim 6, characterized in that, The C18 column can be used alone or in series.

8. The method for detecting epinephrine sulfonate impurities in adrenaline-based drugs according to claim 4, characterized in that, The range of detectors that can be selected in setting the chromatographic conditions includes: electrochemical detectors and fluorescence detectors.

9. The method for detecting epinephrine sulfonate impurities in adrenaline-based drugs according to claim 4, characterized in that, The detector used in setting the chromatographic conditions is an electrochemical detector, preferably an amperometric detector.

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