Method for detecting acetaldehyde content of isavuconazole sulfate for injection

Abstract

The invention discloses a method for detecting acetaldehyde content of isavuconazole sulfate for injection, and belongs to the technical field of pharmaceutical analysis. The detection method comprises the following steps: (1) reacting a test sample with a derivative reagent to obtain a test sample solution; and (2) injecting the test solution into a liquid chromatograph for detection. According to the method, acetaldehyde reacts with a derivatization reagent under an acidic condition, ultraviolet absorption and chromatographic retention of acetaldehyde are enhanced, the derivatization reagent is regulated to be 2-nitrophenylhydrazine, a Syncronis C18 chromatographic column with the particle size of 3 microns is used, and a blank solution does not interfere with detection of acetaldehyde derivatives during detection. In addition, the detection method has high sensitivity and specificity and is easy to operate, the separation degree meets the standard, and the blank of content detection of the isavuconazole sulfate acetaldehyde for injection is filled.

Description

Technical Field

[0001] The present application relates to a method for detecting the acetaldehyde content of isavuconazole sulfate for injection, belonging to the technical field of drug analysis. Background Art

[0002] Isavuconazole sulfate is a triazole antifungal drug used to treat serious infections caused by specific fungi in adult patients. Isavuconazole sulfate is a prodrug of isavuconazole, which is converted into its active form, isavuconazole, in the body to exert its antifungal effect.

[0003] Acetaldehyde is a degradation product of isavuconazole sulfate. The degradation product acetaldehyde is related to clinically found side effects such as increased alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase. The produced acetaldehyde will pose a potential risk to the patient's medication safety. Therefore, it is necessary to control the residual amount of acetaldehyde and establish corresponding methods to control acetaldehyde. However, there is no report on the detection of acetaldehyde content in isavuconazole sulfate injection. Summary of the invention

[0004] According to one aspect of the present application, a method for detecting the acetaldehyde content of isavuconazole sulfate for injection is provided, wherein acetaldehyde reacts with a derivatization reagent under acidic conditions to enhance the ultraviolet absorption and chromatographic retention of acetaldehyde, the derivatization reagent is regulated to be 2-nitrophenylhydrazine, and a Syncronis C18 chromatographic column with a particle size of 3 μm is used, and the blank solution does not interfere with the detection of acetaldehyde derivatives during detection. In addition, the detection method of the present application has high sensitivity and specificity, is simple to operate, and the separation meets the standard, filling the gap in the detection of acetaldehyde content of isavuconazole sulfate for injection.

[0005] A method for detecting the acetaldehyde content of isavuconazole sulfate for injection, comprising:

[0006] (1) After the test sample and the derivatization reagent react, a test sample solution is obtained;

[0007] (2) Inject the test solution into the liquid chromatograph for detection.

[0008] Optionally, the derivatization agent is selected from any one or more of 2,4-dinitrophenylhydrazine, 2-nitrophenylhydrazine, and 2,4-dinitrophenol.

[0009] Optionally, the mass ratio of isavuconazole sulfate to the derivatization reagent in the test sample is (1-2):1; preferably, the mass ratio of isavuconazole sulfate to the derivatization reagent in the test sample is 1.5:1.

[0010] Optionally, the solvent of the test solution is an aqueous solution of acetonitrile.

[0011] Optionally, the volume ratio of acetonitrile to water in the aqueous solution of acetonitrile is (0.8-1.2):1; preferably, the volume ratio of acetonitrile to water in the aqueous solution of acetonitrile is 1:1.

[0012] Optionally, the concentration of the test solution is (1.5-2.5) mg / mL.

[0013] Alternatively, the test solution is prepared as follows:

[0014] (1) 2-nitrophenylhydrazine solution: 600 mg of 2-nitrophenylhydrazine (30 wt% water-moistened) is added with 40 ml of acetonitrile and 5 ml of hydrochloric acid, placed in a 50 ml volumetric flask, dissolved by ultrasonication, diluted to the mark with water, and shaken to obtain;

[0015] (2) Preparation of test sample stock solution: Take one bottle of this product, add an appropriate amount of solvent to dissolve it, transfer all of it to a 50 ml volumetric flask, rinse the vial several times, add all of the washing solution to the volumetric flask, dilute to the scale with solvent, shake well, and the solution is ready.

[0016] (3) Preparation of test solution: Accurately measure 5 ml of the test sample stock solution and place it in a 20 ml volumetric flask. Accurately add 3 ml of 2-nitrophenylhydrazine solution and dilute to the mark with solvent. Shake well to obtain the solution.

[0017] Optionally, the chromatographic conditions include:

[0018] Chromatographic column: octadecylsilane bonded silica gel chromatographic column;

[0019] Mobile phase: water-acetonitrile-trifluoroacetic acid.

[0020] Optionally, the volume ratio of water, acetonitrile and trifluoroacetic acid in the mobile phase is (45-55):(55-45):0.1.

[0021] Optionally, the chromatographic column conditions further include one or more of the following conditions:

[0022] (a1) Column length is 100-250 mm;

[0023] (a2) diameter 2.1-4.6 mm;

[0024] (a3) Particle size is 1.7-3μm.

[0025] Optionally, the chromatographic column is Syncronis C18, with specifications of 4.6×150 mm, 3 μm.

[0026] Optionally, the chromatographic conditions also include: a flow rate of 0.4-0.6 mL / min; preferably, the flow rate is 0.5 mL / min.

[0027] Optionally, the chromatographic conditions further include one or more of the following conditions:

[0028] (b1) Column temperature is 25-35°C;

[0029] (b2) detection wavelength is 250-400nm;

[0030] (b3) The injection volume is 8-12 μl.

[0031] The beneficial effects of this application include:

[0032] 1. The ultraviolet absorption of acetaldehyde is weak. The present application utilizes acetaldehyde to react with a derivatization agent under acidic conditions to enhance the ultraviolet absorption and chromatographic retention of acetaldehyde. The inventors found in the process of studying the acetaldehyde content of isavuconazole sulfate for injection that the blank solution would interfere with the determination of acetaldehyde derivatives after testing using the conventional derivatization agent 2,4-dinitrophenylhydrazine used by those skilled in the art. The general method in the prior art is to solve the interference of the blank solution on the acetaldehyde derivatives by crystallizing and purifying the derivatization agent, but this increases the complexity of the detection steps. The inventors found in the experiment that when the derivatization agent is 2-nitrophenylhydrazine and the chromatographic column is replaced with a Syncronis C18 chromatographic column with a particle size of 3 μm, the pretreatment method of crystallizing and purifying the derivatization agent can solve the problem of the interference of the blank solution on the acetaldehyde derivatives caused by the conventional derivatization agent, which is a significant improvement over the prior art.

[0033] 2. This application uses 2-nitrophenylhydrazine as a derivatization reagent and Syncronis C18 with a particle size of 3 μm as a chromatographic column. The inventors found that the response of the reference substance was low and the sensitivity could not meet the requirements when performing acetaldehyde detection. After a series of studies, the inventors found that when the flow rate was 0.4-0.6 ml / min, the retention time of the acetaldehyde derivative was 7.609 min, the S / N was 413, and the quantitative limit could reach 3% of the limit concentration, which could meet the sensitivity requirements, and the blank solution did not interfere with the determination of the acetaldehyde derivative.

[0034] 3. Those skilled in the art generally increase the recovery rate of spiked test sample acetaldehyde by increasing the injection volume, and the injection volume is generally higher than 15 μL. In the present application, the concentration of the test sample solution is controlled to be (1.5-2.5) mg / mL, and the injection volume is 10 μL, which can make the recovery rate of acetaldehyde between 98.5% and 100.3%. The increase in the concentration of the test sample may reduce the recovery rate of acetaldehyde due to the easy encapsulation of acetaldehyde molecules, and reducing the solubility of the test sample will also cause the acetaldehyde recovery rate to fail to meet the requirements.

[0035] 4. The method for detecting the acetaldehyde content of isavuconazole sulfate for injection provided in the present application has high sensitivity and specificity, is simple to operate, and the separation degree meets the standards, thus filling the gap in the detection of acetaldehyde content of isavuconazole sulfate for injection. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 The HPLC chromatograms of the reference solution and blank solution of Example 1 are shown in FIG.

[0037] Figure 2 The HPLC chromatograms of the reference solution and the blank solution of Example 4 are shown in FIG.

[0038] Figure 3 The HPLC chromatogram and peak result graph of the reference solution of Example 5 are shown;

[0039] Figure 4 This is the linear graph of acetaldehyde limit. DETAILED DESCRIPTION

[0040] The present application is described in detail below with reference to embodiments, but the present application is not limited to these embodiments.

[0041] Unless otherwise specified, the raw materials in the examples of this application were purchased through commercial channels.

[0042] Example 1

[0043] Solution preparation:

[0044] 2,4-Dinitrophenylhydrazine solution: 200 mg of 2,4-dinitrophenylhydrazine is placed in a 50 ml volumetric flask, and 40 ml of acetonitrile and 3 ml of hydrochloric acid are added. Dissolve by ultrasonication, dilute to the mark with acetonitrile, and shake well to obtain;

[0045] Blank solution: Accurately measure 1 ml of 2,4-dinitrophenylhydrazine solution, place it in a 20 ml volumetric flask, dilute to the scale with solvent, shake well, and obtain.

[0046] Reference solution: Take 26.7 mg of acetaldehyde reference substance, accurately weigh it, place it in a 100 ml volumetric flask, add solvent to dissolve and dilute to the scale, shake well, accurately measure 1 ml, place it in a 20 ml volumetric flask, add 1 ml of 2,4-dinitrophenylhydrazine solution, dilute to the scale with solvent, shake well, and obtain.

[0047] The chromatographic conditions were as follows:

[0048] Chromatographic column: Waters Symmetry C18 (4.6×150mm, 3.5μm)

[0049] Column temperature: 30°C

[0050] Flow rate: 1.0ml / min

[0051] Detection wavelength: 360nm

[0052] Injection volume: 10 μl

[0053] Mobile phase: water-acetonitrile-trifluoroacetic acid (volume ratio 50:50:0.1)

[0054] Solvent: 50Vol% acetonitrile

[0055] Running time: 20 minutes

[0056] The HPLC chromatograms of the reference solution and blank solution are shown in Figure 1 The solution with the largest peak fluctuation is the reference solution. Figure 1 It can be seen that the blank solution will interfere with the determination of acetaldehyde derivatives, and the blank interference accounts for 0.9% of the peak area of ​​the reference solution.

[0057] Example 2

[0058] Solution preparation: 2-nitrophenylhydrazine solution: 2-nitrophenylhydrazine (30wt% water-moistened) 200mg, put into a 50ml volumetric flask, add 40ml acetonitrile and 3ml hydrochloric acid, sonicate to dissolve, dilute to scale with water, shake well, and obtain;

[0059] Blank solution: Accurately measure 1 ml of 2-nitrophenylhydrazine solution, place it in a 20 ml volumetric flask, dilute to the scale with solvent, shake well, and obtain.

[0060] Reference solution: Take 26.7 mg of acetaldehyde reference substance, accurately weigh it, place it in a 100 ml volumetric flask, add solvent to dissolve and dilute to the scale, shake well, accurately measure 1 ml, place it in a 20 ml volumetric flask, add 1 ml of 2-nitrophenylhydrazine solution, dilute to the scale with solvent, shake well, and obtain.

[0061] The chromatographic conditions were the same as in Example 1.

[0062] Under this condition, the blank solution still interferes with the determination of acetaldehyde derivatives, and the blank interference accounts for 0.3% of the peak area of ​​the reference solution.

[0063] Example 3

[0064] The solution was prepared as in Example 2.

[0065] The chromatographic conditions were as follows:

[0066] Chromatographic column: ZORBAX Eclipse Plus C18 (4.6×150mm, 1.8μm)

[0067] Column temperature: 30°C

[0068] Flow rate: 1.0ml / min

[0069] Detection wavelength: 360nm

[0070] Injection volume: 10 μl

[0071] Mobile phase: water-acetonitrile-trifluoroacetic acid (volume ratio 50:50:0.1)

[0072] Solvent: 50% acetonitrile

[0073] Running time: 20 minutes

[0074] Under this condition, the blank solution will still interfere with the determination of acetaldehyde derivatives.

[0075] Example 4

[0076] Solution preparation: 2-nitrophenylhydrazine solution: 2-nitrophenylhydrazine (30wt% water-moistened) 200mg, put into a 50ml volumetric flask, add 40ml acetonitrile and 3ml hydrochloric acid, sonicate to dissolve, dilute to scale with water, shake well, and obtain;

[0077] Blank solution: Accurately measure 1 ml of 2-nitrophenylhydrazine solution, place it in a 20 ml volumetric flask, dilute to the scale with solvent, shake well, and obtain.

[0078] Reference solution: Take 26.7 mg of acetaldehyde reference substance, accurately weigh it, place it in a 100 ml volumetric flask, add solvent to dissolve and dilute to the scale, shake well, accurately measure 1 ml, place it in a 20 ml volumetric flask, add 1 ml of 2-nitrophenylhydrazine solution, dilute to the scale with solvent, shake well, and obtain.

[0079] The chromatographic conditions were as follows:

[0080] Chromatographic column: Syncronis C18 (4.6×150mm, 3μm)

[0081] Column temperature: 30°C

[0082] Flow rate: 1.0ml / min

[0083] Detection wavelength: 360nm

[0084] Injection volume: 10 μl

[0085] Mobile phase: water-acetonitrile-trifluoroacetic acid (volume ratio 50:50:0.1)

[0086] Solvent: 50Vol% acetonitrile

[0087] Running time: 20 minutes

[0088] The HPLC chromatograms of the reference solution and blank solution are shown in Figure 2 The reference solution has the largest peak fluctuation. Figure 2 It can be seen that under this condition, the blank solution does not interfere with the determination of acetaldehyde derivatives. However, the response of the reference substance is low, with an S / N of 72, and the sensitivity does not meet the requirements.

[0089] Example 5

[0090] The chromatographic conditions were the same as those in Example 4, except that the flow rate was 0.5 ml / min.

[0091] Solution preparation: 2-nitrophenylhydrazine solution: 600 mg of 2-nitrophenylhydrazine (30 wt% water-moistened) is placed in a 50 ml volumetric flask, and 40 ml of acetonitrile and 5 ml of hydrochloric acid are added. Dissolve by ultrasonication, dilute to the mark with water, and shake well to obtain;

[0092] Blank solution: Accurately measure 3 ml of 2-nitrophenylhydrazine solution, place it in a 20 ml volumetric flask, dilute to the scale with solvent, shake well, and obtain.

[0093] Reference solution: Take 53.5 mg of acetaldehyde reference substance, accurately weigh it, place it in a 100 ml volumetric flask, add solvent to dissolve and dilute to the scale, shake well, accurately measure 1 ml, place it in a 20 ml volumetric flask, add 3 ml of 2-nitrophenylhydrazine solution, dilute to the scale with solvent, shake well, and obtain.

[0094] The HPLC chromatogram and peak results of the reference solution are shown in Figure 3 .

[0095] In the chromatogram of the reference solution, the retention time of the acetaldehyde derivatives is 7.609 min, the S / N is 413, the quantitative limit can reach 3% of the limit concentration, which can meet the sensitivity requirement, and the blank solution does not interfere with the determination of the acetaldehyde derivatives.

[0096] Methodological validation:

[0097] (1) Specificity and injection precision

[0098] Solvent: Measure 500ml of water and 500ml of acetonitrile, mix well, and you have it;

[0099] 2-Nitrophenylhydrazine solution: 600 mg of 2-nitrophenylhydrazine (30 wt% water-moistened) is added with 40 ml of acetonitrile and 5 ml of hydrochloric acid, placed in a 50 ml volumetric flask, dissolved by ultrasonic, diluted to the mark with water, and shaken to obtain;

[0100] Blank solution: Measure 3 ml of 2-nitrophenylhydrazine solution, place it in a 20 ml volumetric flask, dilute to the scale with solvent, shake well, and obtain.

[0101] Blank excipient solution: Take 10 mg of mannitol, place it in a 20 ml volumetric flask, add an appropriate amount of solvent to dissolve it, then accurately add 3 ml of 2-nitrophenylhydrazine solution, dilute to the scale with solvent, shake well, and obtain.

[0102] Reference substance stock solution: Take 53.5 mg of acetaldehyde reference substance, weigh accurately, place in a 100 ml volumetric flask, add solvent to dissolve and dilute to scale, shake well, and obtain;

[0103] Reference solution: Accurately measure 1 ml and place in a 20 ml volumetric flask, accurately add 3 ml of 2-nitrophenylhydrazine solution, and dilute to the scale with solvent, shake well, and obtain.

[0104] Stock solution of test sample: Take 1 bottle of this product, add appropriate amount of solvent to dissolve it, transfer all to a 50ml volumetric flask, rinse the vial several times, add all the washing solution into the volumetric flask, dilute to the scale with solvent, shake well, and you have it.

[0105] Test solution: Accurately measure 5 ml of the test sample stock solution, place it in a 20 ml volumetric flask, accurately add 3 ml of 2-nitrophenylhydrazine solution, and dilute to the scale with solvent, shake well, and obtain.

[0106] Accurately measure 10 μl of blank solution, blank excipient solution, reference solution, and test solution, respectively, inject into the liquid chromatograph, and record the chromatogram. The results are shown in the table below:

[0107] Injection precision results

[0108] name 1 2 3 4 5 RSD% Peak area 189.915 190.319 190.178 190.478 190.530 0.2

[0109] From the results in the table above, we can see that the reference solution was injected 5 times continuously, and the RSD of the peak area of ​​acetaldehyde derivatives was 0.2%, which was less than 5.0%; the blank solution, blank auxiliary material and test solution did not interfere with the detection of acetaldehyde derivatives. The instrument has good injection precision and good method specificity.

[0110] (2) Solution stability

[0111] Blank solvent, reference solution, test solution: same as specificity and injection precision;

[0112] Take the reference solution and the test solution, place them at room temperature and refrigerate at 5℃ for different time periods, accurately measure 10μl of each solution, inject into the liquid chromatograph, and record the chromatogram. The results are shown in the following table.

[0113] Reference solution -5℃

[0114] Time (h) Peak area Compared with 0h RD (%) 0 189.915 / 4.2 191.040 0.3 23.6 192.543 0.7 27.6 192.215 0.6

[0115] Reference solution - room temperature

[0116] Time (h) Peak area Compared with 0h RD (%) 0 182.531 / 5.9 193.181 2.8 23.4 202.541 5.2 28.4 207.846 6.5

[0117] Test solution -5℃

[0118] Time (h) Peak area Compared with 0h RD (%) 0 7.575 / 2.8 11.291 19.7

[0119] From the test results in the above table, we can see that the reference solution is placed at 5°C for 27.6 hours and at room temperature for 28.4 hours. Compared with 0 hour, the RD value at each time point is less than 10.0%. The reference solution is stable at 5°C for 27.6 hours and at room temperature for 28.4 hours. The test solution is placed at 5°C for 2.8 hours. Compared with 0 hour, the RD value of the peak area is 19.7%, which is greater than 10.0%, indicating that the test solution is unstable at 5°C for 2.8 hours and the test solution needs to be freshly prepared before use.

[0120] (3) Linear

[0121] Solvent / Blank Solvent: Same as specificity and injection precision;

[0122] Reference substance stock solution: same as specificity and injection precision;

[0123] The linear solution of acetaldehyde is prepared as shown in the following table.

[0124] Linear preparation of acetaldehyde

[0125]

[0126]

[0127] Take 10 μl of each of the above solutions and inject them, and record the chromatogram. The test results are shown in the following table.

[0128] Acetaldehyde linear

[0129]

[0130] The linear graph of acetaldehyde limit is shown in Figure 4 From the test results, it can be seen that when the concentration of acetaldehyde is in the range of 0.216μg / ml to 21.630μg / ml, the linear equation is y=17.6168x-0.7948, the correlation coefficient is 1.0000, which is greater than 0.9900, and the ratio of the absolute value of the Y-axis intercept to the peak area of ​​100% limit concentration is 0.4%, which is less than 10.0%. The concentration and the peak area show a good linear relationship.

[0131] (4) Limit of quantification and detection limit

[0132] Solvent / Blank Solvent: Same as specificity and injection precision;

[0133] Reference substance stock solution: same as specificity and injection precision;

[0134] Quantitative limit stock solution: accurately measure 2 ml of reference stock solution, place in a 100 ml volumetric flask, dilute to scale with solvent, shake well, and obtain;

[0135] LOQ solution: accurately measure 1 ml of the quantitative limit stock solution, place it in a 20 ml volumetric flask, add 3 ml of 2-nitrophenylhydrazine solution, dilute to the scale with solvent, shake well, and obtain;

[0136] LOD solution: Accurately measure 0.5 ml of the quantitative limit stock solution, place it in a 20 ml volumetric flask, add 3 ml of 2-nitrophenylhydrazine solution, dilute to the scale with solvent, shake well, and obtain.

[0137] Take 10 μl of the quantitative limit solution (six injections in a row) and the detection limit solution respectively, inject them into the liquid chromatograph, and record the chromatogram. The test results are shown in the table below.

[0138] Limit of Quantitation Test (S / N)

[0139] name LOQ-1 LOQ-2 LOQ-3 LOQ-4 LOQ-5 LOQ-6 Acetaldehyde derivatives 30 32 34 22 29 29

[0140] Limit of quantitation test (peak area)

[0141] name LOQ-1 LOQ-2 LOQ-3 LOQ-4 LOQ-5 LOQ-6 RSD(%) Acetaldehyde derivatives 4.053 3.743 3.890 3.957 3.721 4.029 3.7

[0142] Limit of Quantitation Test

[0143] name LOQ concentration (μg / ml) Equivalent to limit concentration (%) Acetaldehyde 0.216 2.0

[0144] Detection limit test

[0145] name LOD concentration (μg / ml) Equivalent to limit concentration (%) S / N Acetaldehyde 0.108 1.0 16

[0146] From the test results, it can be seen that the S / N of the acetaldehyde quantitative limit solution is greater than 10, the concentration is less than 20.0% of the limit concentration, and the RSD value of the peak area of ​​6 consecutive injections of LOQ solution is 3.7%, which is less than 15.0%, indicating that this method can meet the requirements for acetaldehyde quantitative detection; the S / N of the acetaldehyde detection limit solution is greater than 3, and the concentration is less than 10.0% of the limit concentration, which can meet the acetaldehyde detection requirements.

[0147] (5) Accuracy

[0148] Solvent / Blank Solvent: Same as specificity and injection precision;

[0149] Reference substance stock solution: same as specificity and injection precision;

[0150] Reference solution: same as specificity and injection precision;

[0151] Stock solution of test sample: Take 2 bottles of this product, add appropriate amount of solvent to dissolve, transfer all to a 100ml volumetric flask, rinse the vial several times, add all the washing solution into the volumetric flask, dilute to the scale with solvent, shake well, and obtain.

[0152] Test solution: Accurately measure 5 ml of the test sample stock solution, place it in a 20 ml volumetric flask, accurately add 3 ml of 2-nitrophenylhydrazine solution, and dilute to the scale with solvent, shake well, and obtain.

[0153] 50% spiked test solution: accurately measure 5 ml of the test stock solution, place it in a 20 ml volumetric flask, accurately add 0.5 ml of the reference stock solution, then accurately add 3 ml of 2-nitrophenylhydrazine solution, dilute to the mark with solvent, shake well, and prepare 3 copies in parallel.

[0154] 100% spiked test solution: accurately measure 5 ml of the test stock solution, place it in a 20 ml volumetric flask, accurately add 1.0 ml of the reference stock solution, then accurately add 3 ml of 2-nitrophenylhydrazine solution, dilute to the mark with solvent, shake well, and prepare 3 copies in parallel.

[0155] 150% spiked test solution: Accurately measure 5 ml of the test stock solution, place in a 20 ml volumetric flask, accurately add 1.5 ml of the reference stock solution, then accurately add 3 ml of 2-nitrophenylhydrazine solution, dilute to the mark with solvent, shake well, and prepare 3 times in parallel.

[0156] Take 10 μl of each of the above solutions, inject into the liquid chromatograph, and record the chromatogram. The test results are shown in the following table.

[0157] Acetaldehyde recovery results

[0158]

[0159] From the above test results, it can be seen that the recoveries of acetaldehyde in the nine spiked test solutions were all between 98.5% and 100.3%, and within the range of 80.0% to 120.0%, the average recovery was 99.3%, and the RSD value was 0.6%, which was less than 10.0%, meeting the requirements, indicating that the accuracy of this method was good.

[0160] (6) Repeatability

[0161] Reference substance stock solution: Take 53.5 mg of acetaldehyde reference substance, weigh accurately, place in a 100 ml volumetric flask, add solvent to dissolve and dilute to scale, shake well, and obtain;

[0162] Reference solution: Accurately measure 1 ml and place in a 20 ml volumetric flask, accurately add 3 ml of 2-nitrophenylhydrazine solution, and dilute to the scale with solvent, shake well, and obtain.

[0163] Stock solution of test sample: Take 1 bottle of this product, add appropriate amount of solvent to dissolve it, transfer all to a 50ml volumetric flask, rinse the vial several times, add all the washing solution into the volumetric flask, dilute to the scale with solvent, shake well, and you have it.

[0164] Test solution: Accurately measure 5 ml of the test stock solution, place it in a 20 ml volumetric flask, accurately add 3 ml of 2-nitrophenylhydrazine solution, and dilute to the mark with solvent, shake well, and you have it. Prepare fresh before use. (Prepare 6 portions in parallel)

[0165] Take 10 μl of each of the above solutions, inject into the liquid chromatograph, and record the chromatogram. The test results are shown in the following table.

[0166] Acetaldehyde repeatability test results

[0167]

[0168] From the above test results, it can be seen that the acetaldehyde content of 6 test solutions was calculated by the external standard method, and the detection results met the acceptable standards. This method has good repeatability.

[0169] (7) Durability

[0170] Blank solution, reference solution, test solution: same as specificity and injection precision;

[0171] Take 10μl of blank solution, reference solution and test solution respectively, and measure at different column temperatures (30℃±5℃) and different chromatographic column conditions, and record the chromatograms. The test results are shown in the following table.

[0172] Durability test results - test solution

[0173] condition Test results RD% Normal conditions 128 / Column temperature 25℃ 126 Not calculated Column temperature 35℃ 139 Not calculated Chromatographic columns from the same brand but different batches 184 Not calculated

[0174] From the above test results, it can be seen that under the same column temperature (30℃±5℃) and different batches of chromatographic columns, the blank solution does not interfere with the detection of acetaldehyde derivatives; the acetaldehyde content of the test solution is calculated by the external standard method compared with normal conditions, and the detection results all meet the requirements. In summary, this method proves that it has good durability.

[0175] The above are only a few embodiments of the present application and do not constitute any form of limitation to the present application. Although the present application is disclosed as above with preferred embodiments, it is not intended to limit the present application. Any technician familiar with the profession, without departing from the scope of the technical solution of the present application, using the technical content disclosed above to make slight changes or modifications are equivalent to equivalent implementation cases and fall within the scope of the technical solution.

Claims

1. A method for detecting the acetaldehyde content of isavuconazole sulfate for injection, characterized in that: include: (1) After the test sample and the derivatization reagent react, a test sample solution is obtained; (2) Inject the test solution into the liquid chromatograph for detection.

2. The method for detecting the acetaldehyde content of isavuconazole sulfate for injection according to claim 1, characterized in that: The derivatization reagent is selected from any one or more of 2,4-dinitrophenylhydrazine, 2-nitrophenylhydrazine, and 2,4-dinitrophenol.

3. The method for detecting the acetaldehyde content of isavuconazole sulfate for injection according to claim 2, characterized in that: The mass ratio of isavuconazole sulfate and the derivatization reagent in the test sample is (1-2):

1.

4. The method for detecting the acetaldehyde content of isavuconazole sulfate for injection according to claim 1, characterized in that: The solvent of the test solution is an aqueous solution of acetonitrile.

5. The method for detecting the acetaldehyde content of isavuconazole sulfate for injection according to claim 4, characterized in that: The volume ratio of acetonitrile to water in the aqueous solution of acetonitrile is (0.8-1.2):

1.

6. The method for detecting the acetaldehyde content of isavuconazole sulfate for injection according to claim 1, characterized in that: The concentration of the test solution is (1.5-2.5) mg / mL.

7. The method for detecting the acetaldehyde content of isavuconazole sulfate for injection according to any one of claims 1 to 6, characterized in that: Chromatographic conditions included: Chromatographic column: octadecylsilane bonded silica gel chromatographic column; Mobile phase: water-acetonitrile-trifluoroacetic acid.

8. The method for detecting the acetaldehyde content of isavuconazole sulfate for injection according to claim 7, characterized in that: In the mobile phase, the volume ratio of water, acetonitrile and trifluoroacetic acid was (45-55):(55-45):0.

1.

9. The method for detecting the acetaldehyde content of isavuconazole sulfate for injection according to claim 7, characterized in that: The chromatographic column conditions also include one or more of the following conditions: (a1) Column length is 100-250 mm; (a2) diameter 2.1-4.6 mm; (a3) Particle size is 1.7-3μm.

10. The method for detecting the acetaldehyde content of isavuconazole sulfate for injection according to claim 7, characterized in that: Chromatographic conditions also include one or more of the following conditions: (b1) Column temperature is 25-35°C; (b2) detection wavelength is 250-400nm; (b3) The injection volume is 8-12 μl.

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