Method for detecting isopropylideneacetone residue in Nicodil bulk drug
The gas chromatograph combined with the external standard method to detect the residual content of isopropylene acetone in nicordil raw materials has been solved, and the problem of ineffective detection of the existing technology has been achieved, which has achieved rapid, accurate and sensitive detection, ensuring product quality and patient medication safety.
Patent Information
- Application Number
- CN202510120027.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-25
- Publication Date
- 2025-04-29
AI Technical Summary
The prior art cannot effectively detect the residual content of isopropylene acetone in nicordil raw materials, resulting in the inability to meet the detection sensitivity requirements, affecting product quality and patient medication safety.
Using a gas chromatograph combined with an external standard method, the test sample solution and the reference sample solution were prepared, and appropriate chromatographic conditions were set, including DB-1 capillary chromatography column, program temperature rise, inlet temperature, shunt ratio and column flow rate, etc., and the chromatogram was recorded to determine the content of isopropylene acetone.
It realizes rapid, accurate and sensitive detection of isopropylene acetone residues in nicordil raw materials, ensuring product quality and patient medication safety.
Smart Images
Figure CN119901845A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of drug detection and analysis, and in particular to a method for detecting mesityl oxide residues in Nicorandil raw material medicine. Background Art
[0002] Nicorandil, whose chemical name is N-(2-hydroxyethyl) nicotinamide nitrate, is the first ATP-sensitive potassium channel opener used clinically for the treatment of various types of angina pectoris.
[0003]
[0004] Mesityl oxide residues in Nicorandil API are produced by the condensation of two molecules of acetone during the acylation process. Mesityl oxide is highly toxic and similar to acetone and isopropanol in chemical structure. It may have similar toxic effects. Conventional detection methods cannot meet the detection sensitivity requirements. Therefore, the detection of mesityl oxide residual solvents in Nicorandil API is of great quality control significance. Summary of the invention
[0005] Purpose of the invention: In view of the problems existing in the prior art, the present invention provides a method for detecting mesityl oxide residues in nicorandil raw materials, which can effectively realize the detection of mesityl oxide residues in nicorandil raw materials.
[0006] Technical solution: The present invention provides a method for detecting mesityl oxide residues in Nicorandil API, comprising the following steps: S1. Prepare the test solution: accurately weigh the nicorandil sample to be tested, place it in a volumetric flask, add methanol to dissolve and dilute to the scale, and shake well to obtain the test solution; S2. Prepare reference solution: accurately weigh mesityl oxide, place in a volumetric flask, add methanol to dissolve and dilute to scale, shake well to obtain reference solution; S3. Determination: Accurately measure the test solution and the reference solution, inject them into the gas chromatograph respectively, and record the chromatogram; wherein the chromatographic conditions are: Chromatographic column: DB-1 capillary column; Column temperature: Program temperature: start at 60°C, hold for 5 minutes, increase to 230°C at a rate of 40°C per minute, hold for 8 minutes; Injection: injection port temperature was 200°C; split ratio was 10:1; Column flow rate: 4.0 ml per minute; Detector: Hydrogen flame ionization detector (FID), detector temperature is 250℃.
[0007] Further, accurately weigh mesityl oxide, put it in a volumetric bottle, add methanol to dissolve and dilute to the scale, shake well to obtain a reference stock solution; dilute the reference stock solution to obtain a series of mesityl oxide linear solutions with a concentration gradient, and inject and analyze; Linear regression fitting was performed with the linear concentration as the abscissa and the peak area as the ordinate to obtain the external standard method working curve for the determination of mesityl oxide residues in Nicorandil bulk drug.
[0008] Furthermore, mesityl oxide has a good linear relationship in the range of 0.0000405 mg / ml to 0.1215 mg / ml, with a regression equation of y = 1201.5x + 0.7964 and a correlation coefficient of R 2 =0.9996.
[0009] Furthermore, in S1, the concentration of the test solution is 1 g / ml.
[0010] Furthermore, in S2, the concentration of the reference solution is 0.1 mg / ml.
[0011] Preferably, in S3, 1 μl of each of the test solution and the reference solution is accurately measured.
[0012] Beneficial effects: Compared with the prior art, the present invention has the following specific beneficial effects: The detection method provided by the present invention can simply, accurately and quickly detect the content of mesityl oxide, and provides an effective detection method for monitoring the content of mesityl oxide in nicorandil medicine, thereby further ensuring the product quality of nicorandil and the medication safety of patients. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is the chromatogram of mesityl oxide reference solution; Figure 2 is the chromatogram of the test solution; Figure 3 is the standard curve of mesityl oxide. DETAILED DESCRIPTION
[0014] The specific information of the test instruments, reagents and chromatographic columns in the present invention are as follows:
[0015] The present invention is described in detail below in conjunction with the embodiments.
[0016] Implementation 1: The detection method of mesityl oxide residue in Nicorandil API is as follows: Preparation of test solution: Take 1.0 g of the sample to be tested, weigh it accurately, place it in a 10 mL volumetric flask, add methanol and shake to dissolve it, and make up to the mark with methanol.
[0017] Preparation of reference solution: Take an appropriate amount of mesityl oxide, weigh accurately, and quantitatively dilute it with methanol to make a solution containing approximately 0.1 mg per 1 mL. Shake well, seal, and use as the reference solution.
[0018] Detection: Automatic injection, take 1μl of each of the above solutions and inject them into the gas chromatograph, and record the chromatogram. Chromatographic conditions: Use DB-1 (30m×0.530mm, 3.00μm) capillary column as the analytical column; program temperature rise: the starting temperature is 60℃, keep it for 5 minutes, and heat it to 230℃ at a rate of 40℃ per minute, and keep it for 8 minutes; the injection port temperature is 200℃; the split ratio is 10:1; the column flow rate (nitrogen) is 4.0ml per minute; hydrogen flame ionization detector (FID), the detector temperature is 250℃.
[0019] Note: The theoretical plate number calculated based on the mesityl oxide peak should not be less than 5000. If there is a chromatographic peak with the same retention time as the mesityl oxide peak in the chromatogram of the test solution, the mesityl oxide content shall not exceed 0.1% (g / g) according to the peak area calculated by the external standard method.
[0020] The chromatogram of the reference solution is as follows Figure 1 ; Chromatogram of the test solution is as follows Figure 2 .
[0021] Implementation 2: Specificity verification of the detection method of the present invention: (1) Blank solvent: methanol.
[0022] (2) Reference stock solution: Take about 100 mg of mesityl oxide and place it in a 100 ml volumetric flask. Add methanol to dissolve and dilute to the mark. Shake well and use this as the reference stock solution.
[0023] (3) Reference solution: Accurately measure 5 mL of the reference stock solution and place it in a 50 mL volumetric flask. Dilute to the mark with methanol and shake well to prepare the reference solution.
[0024] (4) Test solution: Take about 1.0 g of Nicorandil API, weigh accurately, place in a 10 ml volumetric flask, add methanol and shake to dissolve, then dilute to the mark with methanol as the test solution.
[0025] (5) Test sample spiked solution: Take about 1.0 g of Nicorandil API, weigh accurately, place in a 10 ml volumetric flask, accurately add the reference solution to dissolve it, seal, and shake well to prepare the test sample spiked solution.
[0026] Automatic injection method, take 1 μl of the reference solution, test solution, and test spike solution and inject them into the gas chromatograph to record the chromatogram (chromatographic conditions are the same as in embodiment 1). The specificity results are shown in Table 1.
[0027]
[0028] The results showed that the blank solvent and the test solution had no interference with the determination of mesityl oxide, indicating that the determination method of mesityl oxide residue in Nicorandil API had good specificity.
[0029] Implementation 3: The detection limit and quantitative limit of the detection method of the present invention are: The detection limit and quantification limit were determined by the signal-to-noise ratio method. The quantification limit was determined by the corresponding concentration when the signal-to-noise ratio was 10:1, and the detection limit was determined by the corresponding concentration when the signal-to-noise ratio was 3:1.
[0030] Limit of quantification of mesityl oxide: take about 100 mg of mesityl oxide, accurately weigh, place in a 100 ml volumetric flask, dissolve and dilute to the scale with methanol, shake well, as the mesityl oxide stock solution; accurately measure 5 ml of mesityl oxide stock solution, place in a 50 ml volumetric flask, dilute to the scale with methanol, shake well; accurately measure 1 ml, place in a 10 ml volumetric flask, dilute to the scale with methanol, shake well, accurately measure 1 ml, place in a 20 ml volumetric flask, dilute to the scale with methanol, shake well, as the limit of quantification solution of mesityl oxide, the concentration is 0.50645 μg / ml, which is 0.0051% of the limit.
[0031] Detection limit of mesityl oxide: Accurately measure 1 ml of the control solution, place it in a 10 ml volumetric flask, add methanol to dilute to the scale, shake well; accurately measure 1 ml, place it in a 50 ml volumetric flask, add methanol to dilute to the scale, shake well. This is the detection limit solution of mesityl oxide, with a concentration of 0.20258 μg / ml, which is 0.0020% of the limit.
[0032] Implementation 4: The precision of the detection method of the present invention is: 1. Instrument precision Accurately measure 1µl of the reference solution and repeat the injection 6 times. Calculate the retention time and peak area RSD% of mesityl oxide. The test results are shown in Tables 2 and 3.
[0033]
[0034]
[0035] The results showed that the analytical method for the determination of mesityl oxide residue in Nicorandil had good instrumental precision.
[0036] 2. Repeatability Control stock solution: Take about 100 mg of mesityl oxide and place it in a 100 ml volumetric flask. Add methanol to dissolve and dilute to the scale. Shake well and use this as the control stock solution.
[0037] Reference solution: Accurately measure 5 mL of the reference stock solution and place it in a 50 mL volumetric flask, dilute to the mark with methanol, and shake well to prepare the reference solution.
[0038] Test solution: Take about 1.0 g of Nicorandil raw material, weigh accurately, place in a 10 ml volumetric flask, add methanol and shake to dissolve, and dilute to the mark with methanol as the test solution.
[0039] Test sample spiked solution: Take about 1.0g of Nicorandil API, weigh accurately, place in a 10ml volumetric flask, accurately add reference solution to dissolve, prepare 6 portions in parallel, seal, shake well, and use as test sample spiked solution. Automatically inject 1ul of each of the reference solution, test solution, and test sample spiked solution into the gas chromatograph, and record the chromatogram. The test results are shown in Table 4.
[0040]
[0041] The results showed that the analytical method for the determination of mesityl oxide residue in Nicorandil had good repeatability.
[0042] 3. Intermediate precision Control stock solution: Take about 100 mg of mesityl oxide and place it in a 100 ml volumetric flask. Add methanol to dissolve and dilute to the scale. Shake well and use this as the control stock solution.
[0043] Reference solution: Accurately measure 5 mL of the reference stock solution and place it in a 50 mL volumetric flask, dilute to the mark with methanol, and shake well to prepare the reference solution.
[0044] Test solution: Take about 1.0 g of Nicorandil raw material, weigh accurately, place in a 10 ml volumetric flask, add methanol and shake to dissolve, and dilute to the mark with methanol as the test solution.
[0045] Test sample spike solution: Take about 1.0g of Nicorandil API, weigh accurately, place in a 10ml volumetric flask, accurately add reference solution to dissolve, prepare 6 portions in parallel, seal, shake well, and use as test sample spike solution. Different analysts were tested on different instruments at different times.
[0046] Using automatic injection, 1ul of the reference solution, test solution, and test spike solution were injected into the gas chromatograph and the chromatogram was recorded. The test results are shown in Table 5.
[0047]
[0048] The results showed that the intermediate precision of the analytical method for the determination of mesityl oxide residue in Nicorandil was good.
[0049] Implementation 5: Accuracy (recovery rate) of the detection method of the present invention: Mesityl oxide stock solution: Take about 100 mg of mesityl oxide, weigh accurately, put into a 100 ml volumetric flask, dissolve and dilute to the mark with methanol, shake well, and use this as the mesityl oxide stock solution; Take about 100 mg of mesityl oxide, accurately weigh it, place it in a 100 ml volumetric flask, dissolve it with methanol and dilute it to the mark, shake it well, and use it as the mesityl oxide stock solution; accurately measure 5 ml of the mesityl oxide stock solution, place it in a 50 ml volumetric flask, dilute it to the mark with methanol, shake it well; accurately measure 1 ml, place it in a 10 ml volumetric flask, dilute it to the mark with methanol, shake it well, accurately measure 1 ml, place it in a 20 ml volumetric flask, dilute it to the mark with methanol, shake it well, and use it as the mesityl oxide quantitative limit solution; Reference solution: accurately measure 5 ml of mesityl oxide solution, place it in the same 50 ml volumetric flask, dilute to the mark with methanol, shake well, and use it as the reference solution; Test solution: Take about 1.0 g of Nicorandil API, weigh accurately, place in a 10 ml volumetric flask, dilute with methanol, shake well, and use as the test solution; Quantitative limit recovery solution: Take about 1.0g of Nicorandil API, weigh accurately, place in a 10ml volumetric flask, add the quantitation limit solution, shake well, and prepare three times in parallel as the quantitation limit recovery solution; 80% recovery solution: Take about 1.0g of Nicorandil API, weigh accurately, place in a 10ml volumetric flask, dilute with 80% concentration control solution, shake well, and prepare three times as 80% recovery solution in parallel; 100% recovery solution: Take about 1.0g of Nicorandil API, weigh accurately, place in a 10ml volumetric flask, dilute with 100% concentration control solution, shake well, and prepare three times as 80% recovery solution; 120% recovery solution: Take about 1.0 g of Nicorandil API, weigh accurately, place in a 10 ml volumetric flask, add 120% concentration control solution to dilute, shake well, as the 120% recovery solution, prepare three times in parallel.
[0050] Accurately measure 1 μl of each of the above solutions and inject them into the gas chromatograph to record the chromatogram.
[0051] The test results of mesityl oxide recovery are shown in the following table:
[0052] The results showed that the analytical method for the determination of mesityl oxide residues in Nicorandil had a good recovery rate.
[0053] Implementation 6: Linearity and range of the detection method of the present invention: Stock solution: Take about 100 mg of mesityl oxide and place it in a 100 ml volumetric flask. Add methanol to dissolve and dilute to the scale. Shake well and use this as the control stock solution.
[0054] The stock solution was diluted to prepare a series of concentrations, and linear solutions with a quantitative limit of 120% of the standard limit were prepared. Samples were injected for determination: Preparation of 120% reference solution: 3 mL of the mother solution was accurately pipetted into a 25 mL volumetric flask, diluted to the mark with methanol, and shaken well; Preparation of 100% reference solution: 1 mL of the mother solution was accurately pipetted into a 10 mL volumetric flask, diluted to the mark with methanol, and shaken well; Preparation of 80% reference solution: 2 mL of the mother solution was accurately pipetted into a 2 Prepare 60% reference solution: accurately pipette 3 mL of mother liquor into a 50 mL volumetric flask, dilute to the scale with methanol, shake well, and obtain; prepare 40% reference solution: accurately pipette 2 mL of mother liquor into a 50 mL volumetric flask, dilute to the scale with methanol, shake well, and obtain; prepare 20% reference solution: accurately pipette 1 mL of mother liquor into a 50 mL volumetric flask, dilute to the scale with methanol, shake well, and obtain.
[0055] Automatic injection method, inject 1ul into gas chromatograph, record chromatogram, use linear concentration as horizontal axis and peak area as vertical axis to perform linear regression fitting, and obtain external standard method working curve.
[0056]
[0057] The results show that mesityl oxide is in the range of 0.0000405mg / ml to 0.1215mg / ml, the regression equation is: y = 1201.5x + 0.7964, the correlation coefficient is: R 2 =0.9996, good linear relationship.
[0058] Implementation 7: Durability test of the detection method of the present invention: Control stock solution: Take about 100 mg of mesityl oxide and place it in a 100 ml volumetric flask. Add methanol to dissolve and dilute to the scale. Shake well and use this as the control stock solution.
[0059] Reference solution: Accurately measure 5 mL of the reference stock solution and place it in a 50 mL volumetric flask, dilute to the mark with methanol, and shake well to prepare the reference solution.
[0060] Test solution: Take about 1.0 g of Nicorandil raw material, weigh accurately, place in a 10 ml volumetric flask, add methanol and shake to dissolve, and dilute to the mark with methanol as the test solution.
[0061] Test sample spiked solution: Take about 1.0 g of Nicorandil API, weigh accurately, place in a 10 ml volumetric flask, accurately add reference solution to dissolve it, prepare 6 portions in parallel, seal, shake well, and use as test sample spiked solution.
[0062] Using the automatic injection method, 1ul of each of the reference solution, test solution, and test spiked solution was accurately injected into the gas chromatograph, and the chromatogram was recorded. By changing the column flow rate (4.0±0.2ml / min), injection port temperature (200±10℃), and column temperature (initial temperature 60±5℃), the residual solvent determination method of Nicorandil was used to determine and record the chromatogram. The RSD% of mesityl oxide was calculated.
[0063]
[0064] The results showed that the analytical method for determination of mesityl oxide residue in Nicorandil had good robustness.
[0065] Implementation 8: Solution stability test of the detection method of the present invention: Control stock solution: Take about 100 mg of mesityl oxide and place it in a 100 ml volumetric flask. Add methanol to dissolve and dilute to the scale. Shake well and use this as the control stock solution.
[0066] Reference solution: Accurately measure 5 mL of the reference stock solution and place it in a 50 mL volumetric flask, dilute to the mark with methanol, and shake well to prepare the reference solution.
[0067] Test solution: Take about 1.0 g of Nicorandil raw material, weigh accurately, place in a 10 ml volumetric flask, add methanol and shake to dissolve, and dilute to the mark with methanol as the test solution.
[0068] Test sample spiked solution: Take about 1.0 g of Nicorandil API, weigh accurately, place in a 10 ml volumetric flask, accurately add reference solution to dissolve it, prepare 6 portions in parallel, seal, shake well, and use as test sample spiked solution.
[0069] Take 1 μl of each reference solution, test solution, and test spike solution and inject them into the gas chromatograph to record the chromatogram. Take the reference solution and test spike solution and inject them at 0, 2, 4, 8, 12, and 24 hours for 24 consecutive hours, calculate the RSD of the peak area, and conduct a stability test. Record the chromatogram.
[0070]
[0071] The results showed that the analytical method for the determination of mesityl oxide residue in Nicorandil had good solution stability within 24 hours.
[0072] The above embodiments are only for illustrating the technical concept and features of the present invention, and their purpose is to enable people familiar with the technology to understand the content of the present invention and implement it accordingly, and they cannot be used to limit the protection scope of the present invention. Any equivalent transformation or modification made according to the spirit of the present invention should be included in the protection scope of the present invention.
Claims
1. A method for detecting mesityl oxide residue in Nicorandil API, characterized in that: The following steps are involved: S1. Prepare the test solution: accurately weigh the nicorandil sample to be tested, place it in a volumetric flask, add methanol to dissolve and dilute to the scale, and shake well to obtain the test solution; S2. Prepare reference solution: accurately weigh mesityl oxide, place in a volumetric flask, add methanol to dissolve and dilute to scale, shake well to obtain reference solution; S3. Determination: Accurately measure the test solution and the reference solution, inject them into the gas chromatograph respectively, and record the chromatogram; wherein the chromatographic conditions are: Chromatographic column: DB-1 capillary column; Column temperature: Program temperature: start at 60°C, hold for 5 minutes, increase to 230°C at a rate of 40°C per minute, hold for 8 minutes; Injection: injection port temperature was 200°C; split ratio was 10:1; Column flow rate: 4.0 ml per minute; Detector: Hydrogen flame ionization detector (FID), detector temperature is 250℃.
2. The method for detecting mesityl oxide residue in Nicorandil bulk drug according to claim 1, characterized in that: Accurately weigh mesityl oxide, place in a volumetric flask, add methanol to dissolve and dilute to scale, shake well to obtain a reference stock solution; dilute the reference stock solution to obtain a series of mesityl oxide linear solutions with a concentration gradient, and inject for analysis; Linear regression fitting was performed with the linear concentration as the abscissa and the peak area as the ordinate to obtain the external standard method working curve for the determination of mesityl oxide residues in Nicorandil bulk drug.
3. The method for detecting mesityl oxide residue in Nicorandil bulk drug according to claim 2, characterized in that: Mesityl oxide has a good linear relationship in the range of 0.0000405mg / ml to 0.1215mg / ml. The regression equation is: y = 1201.5x + 0.7964, and the correlation coefficient is: R 2 =0.9996.
4. The method for detecting mesityl oxide residue in Nicorandil bulk drug according to claim 1, characterized in that: In S1, the concentration of the test solution is 1 g / ml.
5. The method for detecting mesityl oxide residue in Nicorandil bulk drug according to claim 1, characterized in that: In S2, the concentration of the reference solution is 0.1 mg / ml.
6. The method for detecting mesityl oxide residue in Nicorandil bulk drug according to claim 1, characterized in that: In S3, accurately measure 1 μl of each of the test solution and the reference solution.