Method for detecting N-nitrosoimino diacetic acid in vortioxetine hydrobromide

Abstract

The invention relates to a method for detecting N-nitrosoimino diacetic acid in vortioxetine hydrobromide, which adopts an ion chromatography method, adopts a chromatographic column taking polystyrene-divinylbenzene copolymer as a filler, and adopts a mixed solution of sodium carbonate and sodium bicarbonate as a leacheate. The method disclosed by the invention is simple, good in specificity, accuracy and sensitivity, and beneficial to control on the content of genotoxic impurities in the hydrobromic acid vortioxetine.

Description

Technical Field

[0001] The invention relates to the field of chemical drug analysis, and in particular to a method for detecting related substances in vortioxetine hydrobromide by using ion chromatography. Background Art

[0002] Vortioxetine hydrobromide is a 5-hydroxytryptamine (5-HT) reuptake inhibitor and an agonist, partial agonist or antagonist of the 5-HT receptor. It was jointly developed by Lundbeck Pharmaceuticals of Denmark and Takeda Pharmaceuticals of Japan for the treatment of depression.

[0003]

[0004] N-nitrosoiminodiacetic acid is a nitroso-type genotoxic impurity. It is necessary to control its content in raw materials and preparations to ensure the quality and safety of the products. However, there is currently no literature or other information disclosing the determination method of N-nitrosoiminodiacetic acid in vortioxetine hydrobromide. Therefore, in order to control the quality of vortioxetine hydrobromide products, it is necessary to develop a method for detecting N-nitrosoiminodiacetic acid in vortioxetine hydrobromide.

[0005] . Summary of the invention

[0006] The present invention provides a method for detecting N-nitrosoiminodiacetic acid in vortioxetine hydrobromide. An ion chromatography method is used, which includes the following chromatographic conditions: Chromatographic column: a chromatographic column filled with polystyrene-divinylbenzene copolymer; Eluent: a mixed solution of sodium carbonate and sodium bicarbonate; Flow rate: 0.5~1.0ml / min; Injection volume: 5~100μl; Detector: Conductivity detector.

[0007] Furthermore, in the chromatographic conditions, the chromatographic column is Metrosep A Supp 19 (250×4 mm, 4.6 μm).

[0008] Furthermore, in the chromatographic conditions, the sodium carbonate concentration in the eluent is 1.0-1.5 g / L, and the sodium bicarbonate concentration is 0.10-0.15 g / L, wherein the ratio of the sodium carbonate concentration to the sodium bicarbonate concentration is 10:1.

[0009] Furthermore, in the chromatographic conditions, the flow rate is 0.5~0.7ml / min.

[0010] Furthermore, in the chromatographic conditions, the injection volume is 25-50 μl.

[0011] Furthermore, the column temperature in the chromatographic conditions is 25°C.

[0012] Furthermore, the detection method further comprises the following steps: (1) Preparation of reference solution: Weigh an appropriate amount of N-nitrosoiminodiacetic acid reference solution accurately, add ultrapure water to dissolve and dilute to obtain the reference solution; (2) Preparation of test solution: Weigh an appropriate amount of vortioxetine hydrobromide, add ultrapure water, sonicate, and filter to obtain the test solution; (3) Preparation of eluent: Weigh appropriate amounts of anhydrous sodium carbonate and sodium bicarbonate, add to ultrapure water, mix thoroughly, and filter to obtain the eluent; (4) Detection: Take the above-mentioned reference solution, test solution and ultrapure water and inject them into the ion chromatograph for measurement, and record the chromatogram.

[0013] The method of the present invention adopts an ion chromatograph, selects a chromatographic column with a hydrophilic polystyrene-divinylbenzene copolymer as a filler to measure N-nitrosoiminodiacetic acid in vortioxetine hydrobromide, and can use a reference substance external standard method to calculate the content of N-nitrosoiminodiacetic acid in the test solution. The method of the present invention is simple, has good specificity, accuracy and sensitivity, and has a stable separation effect. It is suitable for the determination of N-nitrosoiminodiacetic acid in vortioxetine hydrobromide and is conducive to the control of the content of genotoxic impurities in vortioxetine hydrobromide. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 is the chromatogram of the reference solution when the chromatographic column is Metrosep A Supp 5 in Example 1; Figure 2 is the chromatogram of the test solution when the chromatographic column is Metrosep A Supp 5 in Example 1; Figure 3 is the chromatogram of blank solvent when the chromatographic column is Metrosep A Supp 5 in Example 1; Figure 4 is the chromatogram of the reference solution when the chromatographic column is Metrosep A Supp 19 in Example 1; Figure 5 is the chromatogram of the test solution when the chromatographic column is Metrosep A Supp 19 in Example 1; Figure 6 is the chromatogram of the spiked test solution when the chromatographic column is Metrosep A Supp 19 in Example 1; Figure 7is the chromatogram of blank solvent when the chromatographic column is Metrosep A Supp 19 in Example 1; Figure 8 is a chromatogram of the reference solution when the eluent is 1.5 g / L sodium carbonate solution (containing 0.15 g / L sodium bicarbonate) in Example 2; Fig. 9 This is the chromatogram of the test solution when the eluent is 1.5 g / L sodium carbonate solution (containing 0.15 g / L sodium bicarbonate) in Example 2. DETAILED DESCRIPTION

[0015] The content of the present invention is further described in detail below by way of embodiments. However, it should not be understood that the above subject matter of the present invention is limited to the following embodiments. In the absence of conflict, the embodiments and features in the embodiments of the present application can be combined with each other at will.

[0016] Example 1 The content of N-nitrosoiminodiacetic acid in vortioxetine hydrobromide was determined by ion chromatography, and the chromatographic conditions were as follows: Chromatographic column: Metrosep A Supp 19 (250×4mm, 4.6μm) or Metrosep A Supp 5 (250×4mm, 5μm); Eluent: 1g / L sodium carbonate solution (containing 0.1g / L sodium bicarbonate); Flow rate: 0.6ml / min; Column temperature: 25°C; Injection volume: 25 μl; Detector: conductivity detector; Regeneration solution: 1% sulfuric acid solution; Collection time: 50min.

[0017] Experimental steps: (1) Preparation of reference solution: Weigh an appropriate amount of N-nitrosoiminodiacetic acid reference substance, accurately measure, add ultrapure water to dissolve and quantitatively dilute to prepare a solution containing about 0.5 μg / ml N-nitrosoiminodiacetic acid in 1 ml; (2) Preparation of test solution: Take about 100 mg of vortioxetine hydrobromide and put it into a 20 ml volumetric flask. Accurately add 10 ml of ultrapure water. After ultrasonication for 10 min, filter with a 0.22 μm filter membrane to obtain the test solution (containing about 10 mg / ml of vortioxetine hydrobromide). (3) Preparation of spiked test solution: Accurately pipette 5 ml of reference solution into a 50 ml volumetric flask, dilute to scale with ultrapure water, shake well, and obtain spiked solution. Take about 100 mg of vortioxetine hydrobromide into a 20 ml volumetric flask, accurately pipette 10 ml of spiked solution, dilute to scale with ultrapure water, shake well, sonicate for 10 min, and filter with a 0.22 μm filter membrane to obtain; (4) Preparation of eluent: Take 1000 mg of anhydrous sodium carbonate and 100 mg of sodium bicarbonate, add them to 1000 ml of ultrapure water, mix thoroughly, and filter with a 0.45 μm filter membrane to obtain the eluent; (5) Detection: Take the above-mentioned reference solution, test solution and ultrapure water (blank solvent) and inject them into the ion chromatograph respectively, measure under the above-mentioned chromatographic conditions, and record the chromatogram. Calculate the content of N-nitrosoiminodiacetic acid in the test solution according to the reference external standard method.

[0018] See the detection spectrum Figure 1 to Figure 7 The results showed that the chromatographic column Metrosep A Supp 19 (250×4mm, 4.6μm) could detect N-nitrosoiminodiacetic acid in vortioxetine hydrobromide, and the method had good specificity.

[0019] Example 2 The content of N-nitrosoiminodiacetic acid in vortioxetine hydrobromide was determined by ion chromatography, and the chromatographic conditions were as follows: Chromatographic column: Metrosep A Supp 19 (250×4 mm, 4.6 μm); Eluent: 1g / L sodium carbonate solution (containing 0.1g / L sodium bicarbonate) or 1.5g / L sodium carbonate solution (containing 0.15g / L sodium bicarbonate); Flow rate: 0.6ml / min; Column temperature: 25°C; Injection volume: 25 μl; Detector: conductivity detector; Regeneration solution: 1% sulfuric acid solution; Collection time: 50min.

[0020] Experimental steps: (1) The preparation of the reference solution and the test solution is the same as in Example 1; (2) Preparation of eluent: Referring to the method for preparing 1 g / L sodium carbonate solution in Example 1, 1 g / L sodium carbonate solution (containing 0.1 g / L sodium bicarbonate) and 1.5 g / L sodium carbonate solution (containing 0.15 g / L sodium bicarbonate) were prepared respectively; (3) Detection: Inject the above-mentioned reference solution, test solution and ultrapure water (blank solvent) into an ion chromatograph respectively, perform the measurement under the above-mentioned chromatographic conditions and record the chromatogram.

[0021] See the detection spectrum Figure 8~Figure 9 The results showed that under the above chromatographic conditions, the determination of N-nitrosoiminodiacetic acid in vortioxetine hydrobromide could be achieved using 1 g / L sodium carbonate solution (containing 0.1 g / L sodium bicarbonate) and 1.5 g / L sodium carbonate solution (containing 0.15 g / L sodium bicarbonate) as eluents.

[0022] Example 3 The content of N-nitrosoiminodiacetic acid in vortioxetine hydrobromide was determined by ion chromatography, and the chromatographic conditions were as follows: Chromatographic column: Metrosep A Supp 19 (250×4 mm, 4.6 μm); Eluent: 1g / L sodium carbonate solution (containing 0.1g / L sodium bicarbonate); Flow rate: 0.5ml / min, 0.6ml / min or 0.7ml / min; Column temperature: 25°C; Injection volume: 25 μl; Detector: conductivity detector; Regeneration solution: 1% sulfuric acid solution; Collection time: 50min.

[0023] Experimental steps: (1) The preparation of reference solution, test solution and eluent is the same as in Example 1; (2) Detection: Inject the above-mentioned reference solution, test solution and ultrapure water (blank solvent) into an ion chromatograph respectively, perform the measurement under the above-mentioned chromatographic conditions and record the chromatogram.

[0024] The test results (see Table 1) show that the flow rate of 0.5 ml / min to 0.7 ml / min can achieve the determination of N-nitrosoiminodiacetic acid in vortioxetine hydrobromide.

[0025] Table 1 .

[0026] Example 4 The content of N-nitrosoiminodiacetic acid in vortioxetine hydrobromide was determined by ion chromatography, and the chromatographic conditions were as follows: Chromatographic column: Metrosep A Supp 19 (250×4 mm, 4.6 μm); Other chromatographic conditions were the same as in Example 1.

[0027] Experimental steps: (1) Preparation of test solution and spiked test solution: Prepare three portions in parallel as in Example 1; (2) Detection: Take the above-mentioned test sample solution and the spiked test sample solution and inject them into the ion chromatograph respectively. Measure them under the above-mentioned chromatographic conditions and record the chromatograms.

[0028] The test results are shown in Table 2. The recovery of N-nitrosoiminodiacetic acid in the three spiked test solutions was 94.2% to 99.3%, the average recovery was 96.8%, and the RSD was 2.7%. The experimental results show that the method of the present invention has good accuracy in determining the content of N-nitrosoiminodiacetic acid in vortioxetine hydrobromide.

[0029] Table 2 .

Claims

1. A method for detecting N-nitrosoiminodiacetic acid in vortioxetine hydrobromide, characterized in that: Ion chromatography was used, including the following chromatographic conditions: Column: Metrosep A Supp 19; Eluent: a mixed solution of sodium carbonate and sodium bicarbonate; Flow rate: 0.5~1.0ml / min; Injection volume: 5~100μl; Detector: Conductivity detector.

2. The method according to claim 1, wherein the sodium carbonate concentration in the eluent is 1.0-1.5 g / L, and the sodium bicarbonate concentration is 0.10-0.15 g / L, wherein: The ratio of sodium carbonate concentration to sodium bicarbonate concentration is 10:

1.

3. The method according to claim 1, wherein the flow rate is 0.5-0.7 ml / min.

4. The method according to claim 1, wherein the injection volume is 25-50 μl. The method according to claim 1 , wherein the column temperature in the chromatographic conditions is 25° C.

6. The method according to any one of claims 1 to 5, further comprising the following steps: (1) Preparation of reference solution: Weigh an appropriate amount of N-nitrosoiminodiacetic acid reference solution accurately, add ultrapure water to dissolve and dilute to obtain the reference solution; (2) Preparation of test solution: Weigh an appropriate amount of vortioxetine hydrobromide, add ultrapure water, sonicate, and filter to obtain the test solution; (3) Preparation of eluent: Weigh appropriate amounts of anhydrous sodium carbonate and sodium bicarbonate, add to ultrapure water, mix thoroughly, and filter to obtain the eluent; (4) Detection: Take the above-mentioned reference solution, test solution and ultrapure water and inject them into the ion chromatograph for measurement, and record the chromatogram.

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