Method for detecting mutagenic impurity desloratadine pyridine nitrogen oxide in desloratadine oral solution

Abstract

The invention relates to the technical field of drug detection, and discloses a method for detecting mutagenic impurity desloratadine pyridine nitrogen oxide in a desloratadine oral solution, which comprises the following specific steps: S1, preparing a test solution: precisely weighing a to-be-detected sample, placing the to-be-detected sample in a measuring flask, adding a mobile phase, dissolving, diluting to a scale, and uniformly shaking to obtain the test solution; s2, preparation of a reference substance solution: precisely weighing a desloratadine pyridine nitrogen oxide reference substance, placing the reference substance in a measuring flask, adding a mobile phase for dissolving and diluting to a scale, and uniformly shaking to obtain the reference substance solution; and S3, determination: precisely measuring the test solution and the reference solution, respectively injecting into a liquid chromatograph, and recording a chromatogram to two times of the retention time of a main component peak. According to the present invention, with the method, the mutagenic impurity desloratadine pyridine nitrogen oxide in the desloratadine oral solution can be accurately and rapidly determined, and the detection method has characteristics of strong specificity and high sensitivity.

Description

Technical Field

[0001] The present invention relates to the technical field of drug detection, and particularly to a method for detecting the mutagenic impurity desloratadine pyridine N-oxide in desloratadine oral solution. Background Art

[0002] Desloratadine, chemically named 8-chloro-6,11-dihydro-11-(4-piperidinylidene)-5H-benzo[5,6]cyclohepta[1,2-b]pyridine, is a non-sedating long-acting tricyclic antihistamine and an active metabolite obtained by deethyl formate in vivo. It takes effect directly without being metabolized by liver CYP3A4 and can relieve the related symptoms of allergic rhinitis or chronic idiopathic urticaria by selectively antagonizing peripheral H1 receptors.

[0003] In desloratadine oral solution, desloratadine is oxidized to produce desloratadine pyridine N-oxide, which is a mutagenic impurity with a warning structure:

[0004]

[0005] The concentration of desloratadine oral solution is 0.5 mg / ml, and the maximum daily dose is 10 ml. Then the maximum daily dose of desloratadine is 5 mg. There is no clear toxicological data for desloratadine pyridine N-oxide. According to the TTC method, desloratadine pyridine N-oxide is 1.5 μg / day, and the limit of desloratadine pyridine N-oxide in desloratadine oral solution is 1.5 μg / 5 mg = 0.03%.

[0006] To ensure the safety and reliability of drugs, an effective method for detecting the mutagenic impurity desloratadine pyridine N-oxide in desloratadine oral solution is needed. Summary of the Invention

[0007] Object of the Invention: Aiming at the problems existing in the prior art, the present invention provides a method for detecting the mutagenic impurity desloratadine pyridine N-oxide in desloratadine oral solution, which can accurately and quickly determine the mutagenic impurity desloratadine pyridine N-oxide in desloratadine oral solution.

[0008] Technical Solution: The present invention provides a method for detecting the mutagenic impurity desloratadine pyridine N-oxide in desloratadine oral solution, comprising the following steps:

[0009] S1. Prepare a test solution: Weigh accurately the test sample to be measured, place it in a volumetric flask, dissolve it with the mobile phase and dilute to the scale, and shake well to obtain the test solution;

[0010] S2. Prepare the reference solution: Weigh accurately the desloratadine pyridine N-oxide reference substance, place it in a volumetric flask, dissolve it with the mobile phase and dilute to the mark, and shake well to obtain the reference solution;

[0011] S3. Determination: Accurately measure the test solution and the reference solution, and inject them into the liquid chromatograph respectively, and record the chromatogram until twice the retention time of the main component peak; among them, the chromatographic conditions are as follows:

[0012] Chromatographic column: Octadecylsilane chemically bonded silica gel;

[0013] Column temperature: 35 °C;

[0014] Injection: Injection volume is 100 μl;

[0015] Mobile phase: Mobile phase A: Aqueous solution of sodium dodecyl sulfate with a concentration of 0.003 mol / L; Mobile phase B: Acetonitrile; The volume ratio of mobile phase A to mobile phase B is 60:40;

[0016] Flow rate: 4.0 ml per minute;

[0017] Detection wavelength: 280 nm.

[0018] Furthermore, the structural formula of the desloratadine pyridine N-oxide is as follows:

[0019]

[0020] Furthermore, in S1, the concentration of the test solution is 0.2 mg / ml.

[0021] Furthermore, in S2, the concentration of the reference solution is 0.06 μg / ml.

[0022] Furthermore, in S3, the specific preparation method of the aqueous solution of sodium dodecyl sulfate is: Take 0.9 g of sodium dodecyl sulfate with a purity of 99% and 0.5 ml of trifluoroacetic acid, mix them, dissolve with water and dilute to 1000 ml.

[0023] Furthermore, if there is a chromatographic peak in the chromatogram of the test solution with the same retention time as that of desloratadine pyridine N-oxide, calculate according to the external standard method with the peak area to obtain the content of desloratadine pyridine N-oxide in the test solution.

[0024] Beneficial effects: Compared with the prior art, the specific beneficial effects of the present invention are as follows:

[0025] The present invention can accurately and rapidly determine the mutagenic impurity of desloratadine pyridine N-oxide in desloratadine oral solution, and the detection method has strong specificity and high sensitivity. The present invention can ensure the stable, safe and controllable quality of the product, improve the drug quality and drug safety of desloratadine oral solution, and provide an effective method for monitoring the content of the mutagenic impurity of desloratadine pyridine N-oxide in desloratadine oral solution. Description of the Drawings

[0026] Figure 1 It is the chromatogram of the blank solvent of the present invention;

[0027] Figure 2 It is the chromatogram of the blank excipient solution of the present invention;

[0028] Figure 3 It is the chromatogram of the reference solution of the present invention;

[0029] Figure 4 It is the chromatogram of the test solution of the present invention;

[0030] Figure 5 It is the chromatogram of the spiked test solution of the present invention;

[0031] Figure 6 It is the linear graph of desloratadine pyridine N-oxide of the present invention. Detailed Embodiments

[0032] The present invention will be introduced in detail below in combination with the embodiments.

[0033] Embodiment 1:

[0034] The specific information of the instruments, reagents and chromatographic columns used in the tests of the present invention is as follows:

[0035] 1. Information on Instruments and Chromatographic Columns

[0036]

[0037] 2. Information on Reagents

[0038] Reagent Name Manufacturer Batch Number Grade / Content Sodium Dodecyl Sulfate Sians 500+S75101MCW1 99％ Trifluoroacetic Acid TEDIA 19100409 HPLC Acetonitrile Fortum DW42A121401 HPLC

[0039] 3. Information on Reference Substances

[0040] Reference Standard Name Manufacturer Batch Number Content Desloratadine Pyridine N-Oxide QCC 07-JAN-23-23 94.66％

[0041] Embodiment 1:

[0042] The detection method for the mutagenic impurity of desloratadine pyridine N-oxide in desloratadine oral solution is as follows:

[0043] Preparation of test solution: Accurately measure an appropriate amount of this product, dissolve it with the mobile phase and quantitatively dilute it to prepare a solution containing about 0.2 mg of desloratadine per 1 ml.

[0044] Preparation of reference solution: Take an appropriate amount of desloratadine pyridine N - oxide reference substance, accurately weigh it, dissolve it with the mobile phase and quantitatively dilute it to prepare a solution containing about 0.06 μg per 1 ml.

[0045] Detection: Accurately measure the test solution and the reference solution, and inject them into the liquid chromatograph respectively. Record the chromatogram of the test solution until 2 times the retention time of the main component peak. If there is a chromatographic peak in the chromatogram of the test solution with the same retention time as that of desloratadine pyridine N - oxide, calculate the content of desloratadine pyridine N - oxide in the test solution by the external standard method based on the peak area. Among them, the chromatographic conditions are as follows: Use octadecylsilane chemically bonded silica gel as the filler (4.6 mm×250 mm, 4 μm); Use an aqueous solution of 0.003 mol / L sodium dodecyl sulfate (take 0.9 g of sodium dodecyl sulfate (purity 99%), 0.5 ml of trifluoroacetic acid, dissolve and dilute to 1000 ml with water) - acetonitrile (60:40) as the mobile phase; The flow rate is 1.0 ml per minute; The detection wavelength is 280 nm; The column temperature is 35°C; The injection volume is 100 μl.

[0046] Embodiment 2:

[0047] Specificity verification of the detection method of the present invention:

[0048] Blank solvent: Mobile phase.

[0049] Blank excipient solution: Accurately measure 4 ml of blank excipients, place them in a 10 - ml volumetric flask, dissolve them with the mobile phase and dilute to the scale, and shake well.

[0050] Test solution: Accurately measure 4 ml of this product, place it in a 10 - ml volumetric flask, dissolve it with the mobile phase and dilute to the scale, and shake well.

[0051] Reference solution: Take an appropriate amount of desloratadine pyridine N - oxide reference substance, accurately weigh it, dissolve it with the mobile phase and quantitatively dilute it to prepare a solution containing about 0.06 μg per 1 ml, and shake well.

[0052] Accurately measure the above - mentioned solutions and inject them into the liquid chromatograph respectively, and record the chromatograms. Examine whether there is interference from the blank solvent and the blank excipient solution on the determination of desloratadine pyridine N - oxide.

[0053] Specificity determination results

[0054]

[0055] The results showed that the blank solvent and blank excipient solution had no interference on the determination of desloratadine pyridine N-oxide, and the resolution between the desloratadine pyridine N-oxide peak and adjacent impurity peaks was good, indicating that the analytical method had good specificity.

[0056] Embodiment 3:

[0057] The precision of the detection method of the present invention:

[0058] 1. Instrument precision:

[0059] Take the reference solution and repeat the determination 6 times according to the desloratadine pyridine N-oxide analysis method of the present invention, and calculate the RSD of the peak area and retention time of desloratadine pyridine N-oxide.

[0060] Instrument precision determination results (JQ1322)

[0061] — Retention Time (min) Peak Area 1 10.744 5269 2 10.740 5380 3 10.739 5559 4 10.752 5210 5 10.752 5330 6 10.749 5413 Average Value 10.746 5360 RSD (%) 0.05 2.28

[0062] Instrument precision determination results (JQ1312)

[0063] — Retention Time (min) Peak Area 1 11.038 5361 2 11.045 5489 3 11.054 5356 4 11.058 5567 5 11.067 5401 6 11.073 5445 Average Value 11.056 5437 RSD (%) 0.12 1.50

[0064] The results showed that the instrument precision of the analytical method was good.

[0065] 2. Repeatability:

[0066] Reference stock solution: Take an appropriate amount of desloratadine pyridine N-oxide reference substance, weigh it accurately, dissolve it with the mobile phase and quantitatively dilute it to a solution containing about 0.6 μg per 1 ml, and shake well.

[0067] Reference solution: Accurately measure 2 ml of the reference stock solution, place it in a 20-ml volumetric flask, dilute it to the mark with the mobile phase, and shake well.

[0068] Test solution: Accurately measure 4 ml of this product, place it in a 10-ml volumetric flask, dissolve it with the mobile phase and dilute it to the mark, and shake well.

[0069] Spiked test solution: Accurately measure 4 ml of this product, place it in a 10-ml volumetric flask, accurately add 1 ml of the reference stock solution, dissolve it with the mobile phase and dilute it to the mark, and shake well (prepare 6 parallel portions).

[0070] According to the desloratadine pyridine N-oxide analysis method, determine 6 portions of the spiked test solution. Examine the recovery rate and RSD of desloratadine pyridine N-oxide in the 6 portions of the spiked test solution.

[0071] Repeatability determination results

[0072]

[0073]

[0074] The results showed that the analysis method had good repeatability.

[0075] 3. Intermediate precision:

[0076] Stock solution of reference substance: Weigh an appropriate amount of desloratadine pyridine N-oxide reference substance accurately, dissolve it in the mobile phase and quantitatively dilute it to prepare a solution containing about 0.6 μg per 1 ml, and shake well.

[0077] Reference substance solution: Accurately measure 2 ml of the stock solution of reference substance, place it in a 20-ml volumetric flask, dilute it to the mark with the mobile phase, and shake well.

[0078] Test solution: Accurately measure 4 ml of this product, place it in a 10-ml volumetric flask, dissolve it with the mobile phase and dilute it to the mark, and shake well.

[0079] Spiked test solution: Accurately measure 4 ml of this product, place it in a 10-ml volumetric flask, accurately add 1 ml of the stock solution of reference substance, dissolve it with the mobile phase and dilute it to the mark, and shake well (prepared in parallel for 12 portions, including 6 portions under repeatability).

[0080] According to the analysis method of desloratadine pyridine N-oxide, determine 12 portions of the spiked test solution. Examine the recovery rate and RSD of desloratadine pyridine N-oxide in the 12 portions of the spiked test solution.

[0081] Results of intermediate precision determination

[0082]

[0083] The results showed that the analysis method had good intermediate precision.

[0084] Embodiment 4:

[0085] Detection limit and quantitation limit of the detection method of the present invention:

[0086] The signal-to-noise ratio method was used to determine the detection limit and quantitation limit. The corresponding concentration at a signal-to-noise ratio of about 3:1 was used as the detection limit, and the corresponding concentration at a signal-to-noise ratio of about 10:1 was used as the quantitation limit.

[0087] (1) Detection limit

[0088] Take an appropriate amount of desloratadine pyridine N-oxide reference substance, weigh it accurately, dissolve it with the mobile phase and quantitatively dilute it to prepare a solution containing about 0.6 μg per 1 ml, shake well, and use it as the stock solution of desloratadine pyridine N-oxide reference substance; accurately measure 0.2 ml of the stock solution of desloratadine pyridine N-oxide reference substance, place it in a 10-ml volumetric flask, quantitatively dilute it to the scale with the mobile phase, and shake well; accurately measure 3 ml, place it in a 10-ml volumetric flask, dilute it to the scale with the mobile phase, and shake well, and use it as the detection limit solution of desloratadine pyridine N-oxide. The detection limit concentration is 0.0035 μg / ml, which is 6% of the limit.

[0089] (2) Quantitation limit

[0090] Take an appropriate amount of desloratadine pyridine N-oxide reference substance, weigh it accurately, dissolve it with the mobile phase and quantitatively dilute it to prepare a solution containing about 0.6 μg per 1 ml, shake well, and use it as the stock solution of desloratadine pyridine N-oxide reference substance; accurately measure 0.2 ml of the stock solution of desloratadine pyridine N-oxide reference substance, place it in a 10-ml volumetric flask, quantitatively dilute it to the scale with the mobile phase, and shake well; accurately measure 3 ml, place it in a 10-ml volumetric flask, dilute it to the scale with the mobile phase, and shake well, and use it as the quantitation limit solution of desloratadine pyridine N-oxide. The quantitation limit concentration is 0.0118 μg / ml, which is 20% of the limit.

[0091] Take the quantitation limit solution of desloratadine pyridine N-oxide and inject it repeatedly for 6 times to examine the RSD of the peak area.

[0092] Determination results of the quantitation limit solution

[0093] Quantitation Limit Peak Area S / N 1 1174 12.15 2 1077 9.63 3 1045 11.31 4 1034 9.30 5 1073 11.38 6 1051 9.85 Average Value 1076 — RSD (%) 4.73 —

[0094] The results show that the precision of the quantitation limit solution of desloratadine pyridine N-oxide is good.

[0095] Embodiment 5:

[0096] Linearity and range of the detection method of the present invention:

[0097] Stock solution of reference substance: Take an appropriate amount of desloratadine pyridine N-oxide reference substance, weigh it accurately, dissolve it with the mobile phase and quantitatively dilute it to prepare a solution containing about 0.6 μg per 1 ml, and shake well.

[0098] Accurately measure 0.2 ml, 0.5 ml, 0.8 ml, 1.0 ml, 1.5 ml, and 2.0 ml of the stock solution of reference substance respectively, place them in 10-ml volumetric flasks, quantitatively dilute them to the scale with the mobile phase, and shake well to obtain linear solution 1 to linear solution 6.

[0099] Take linear solution 1 to linear solution 6 and the quantitation limit solution, inject them into the liquid chromatograph respectively, record the chromatogram, and calculate the linear regression equation.

[0100] Linear determination results

[0101]

[0102]

[0103] The results showed that desloratadine pyridine N-oxide had a good linear relationship in the concentration range of 0.0118 μg / ml to 0.1179 μg / ml, and the linear equation was y = 93513x - 43.822, with R 2 = 0.9984.

[0104] Embodiment 6:

[0105] Accuracy of the detection method of the present invention:

[0106] Stock solution of reference substance: Take an appropriate amount of desloratadine pyridine N-oxide reference substance, weigh it accurately, dissolve it with the mobile phase and quantitatively dilute it to make a solution containing about 0.6 μg per 1 ml, and shake well.

[0107] Reference substance solution: Accurately measure 2 ml of the stock solution of reference substance, place it in a 20-ml volumetric flask, dilute it to the mark with the mobile phase, and shake well.

[0108] Test solution: Accurately measure 4 ml of this product, place it in a 10-ml volumetric flask, dissolve it with the mobile phase and dilute it to the mark, and shake well.

[0109] LOQ spiked test solution: Accurately measure 4 ml of this product, place it in a 10-ml volumetric flask, accurately add 0.2 ml of the stock solution of reference substance, dissolve it with the mobile phase and dilute it to the mark, and shake well (prepare 3 parallel portions).

[0110] 80% spiked test solution: Accurately measure 4 ml of this product, place it in a 10-ml volumetric flask, accurately add 0.8 ml of the stock solution of reference substance, dissolve it with the mobile phase and dilute it to the mark, and shake well (prepare 3 parallel portions).

[0111] 100% spiked test solution: Accurately measure 4 ml of this product, place it in a 10-ml volumetric flask, accurately add 1 ml of the stock solution of reference substance, dissolve it with the mobile phase and dilute it to the mark, and shake well (prepare 3 parallel portions).

[0112] 120% spiked test solution: Accurately measure 4 ml of this product, place it in a 10-ml volumetric flask, accurately add 1.2 ml of the stock solution of reference substance, dissolve it with the mobile phase and dilute it to the mark, and shake well (prepare 3 parallel portions).

[0113] Accurately measure the above solutions, inject them into the liquid chromatograph respectively, and record the chromatogram. Examine the recovery rate and RSD of desloratadine pyridine N-oxide in the spiked test solutions at each level.

[0114] Accuracy determination results

[0115]

[0116]

[0117] The results show that the accuracy of this analysis method is good.

[0118] Embodiment 7:

[0119] Durability of the detection method of the present invention:

[0120] Stock solution of reference substance: Take an appropriate amount of desloratadine pyridine N - oxide reference substance, weigh it accurately, dissolve it with the mobile phase and quantitatively dilute it to prepare a solution containing about 0.6 μg per 1 ml, and shake well.

[0121] Reference substance solution: Accurately measure 2 ml of the stock solution of reference substance, place it in a 20 - ml volumetric flask, dilute it to the mark with the mobile phase, and shake well.

[0122] Test solution: Accurately measure 4 ml of this product, place it in a 10 - ml volumetric flask, dissolve it with the mobile phase and dilute it to the mark, and shake well.

[0123] Spiked test solution: Accurately measure 4 ml of this product, place it in a 10 - ml volumetric flask, accurately add 1 ml of the stock solution of reference substance, dissolve it with the mobile phase and dilute it to the mark, and shake well.

[0124] According to the analysis method of desloratadine pyridine N - oxide, investigate the recovery rate and RSD of desloratadine pyridine N - oxide in the spiked test solution under different wavelengths (280 nm ± 2 nm), different column temperatures (35 ± 2 °C) and different chromatographic column conditions.

[0125] Durability determination results

[0126]

[0127] The results show that the durability of the analysis method of desloratadine pyridine N - oxide is good.

[0128] Embodiment 8:

[0129] Solution stability of the detection method of the present invention:

[0130] Stock solution of reference substance: Take an appropriate amount of desloratadine pyridine N - oxide reference substance, weigh it accurately, dissolve it with the mobile phase and quantitatively dilute it to prepare a solution containing about 0.6 μg per 1 ml, and shake well.

[0131] Reference substance solution: Accurately measure 2 ml of the stock solution of reference substance, place it in a 20 - ml volumetric flask, dilute it to the mark with the mobile phase, and shake well.

[0132] Test sample spiked solution: Accurately measure 4 ml of this product, place it in a 10-ml volumetric flask, accurately add 1 ml of the reference stock solution, dissolve with the mobile phase and dilute to the scale, and shake well.

[0133] According to the analytical method of desloratadine pyridine N-oxide, examine the RSD of the peak areas of desloratadine pyridine N-oxide in the reference solution and the test sample spiked solution within a certain period of time.

[0134] Determination results of the stability of the test sample spiked solution

[0135]

[0136] Determination results of the stability of the reference solution

[0137]

[0138] The results show that: the reference solution is stable within 24 hours, and desloratadine pyridine N-oxide in the test sample spiked solution is stable within 24 hours.

[0139] The above embodiments are only for explaining the technical concept and characteristics of the present invention, and the purpose is to enable those who are familiar with this technology to understand the content of the present invention and implement it accordingly, and it cannot be used to limit the protection scope of the present invention. Any equivalent transformation or modification made according to the spirit and essence of the present invention should be covered within the protection scope of the present invention.

Claims

1. A method for detecting the mutagenic impurity desloratadine pyridine nitrogen oxide in desloratadine oral solution, characterized in that: The following steps are involved: S1. Prepare the test solution: accurately weigh the sample to be tested, place it in a volumetric flask, add the mobile phase to dissolve and dilute to the scale, and shake well to obtain the test solution; S2. Prepare the reference solution: accurately weigh the desloratadine pyridine nitrogen oxide reference substance, place it in a volumetric flask, add the mobile phase to dissolve and dilute to the scale, and shake well to obtain the reference solution; S3. Determination: Accurately measure the test solution and the reference solution, inject them into the liquid chromatograph respectively, and record the chromatogram to twice the retention time of the main component peak; wherein, the chromatographic conditions are: Chromatographic column: octadecylsilane bonded silica gel; Column temperature: 35°C; Injection: injection volume 100 μl; Mobile phase: Mobile phase A: 0.003 mol / L sodium dodecyl sulfate aqueous solution; Mobile phase B: acetonitrile; The volume ratio of mobile phase A to mobile phase B is 60:40; Flow rate: 4.0 ml per minute; Detection wavelength: 280nm.

2. The method for detecting the mutagenic impurity desloratadine pyridine nitrogen oxide in the desloratadine oral solution according to claim 1, characterized in that: The structural formula of the desloratadine pyridine nitrogen oxide is as follows: 。 3. The method for detecting the mutagenic impurity desloratadine pyridine nitrogen oxide in the desloratadine oral solution according to claim 1, characterized in that: In S1, the concentration of the test solution is 0.2 mg / ml.

4. The method for detecting the mutagenic impurity desloratadine pyridine nitrogen oxide in the desloratadine oral solution according to claim 1, characterized in that: In S2, the concentration of the reference solution is 0.06 μg / ml.

5. The method for detecting the mutagenic impurity desloratadine pyridine nitrogen oxide in the desloratadine oral solution according to claim 1, characterized in that: In S3, the specific preparation method of the sodium dodecyl sulfate aqueous solution is: take 0.9 g of sodium dodecyl sulfate with a purity of 99% and 0.5 ml of trifluoroacetic acid, mix them, add water to dissolve and dilute to 1000 ml.

6. The method for detecting the mutagenic impurity desloratadine pyridine nitrogen oxide in the desloratadine oral solution according to claim 1, characterized in that: If there is a chromatographic peak in the chromatogram of the test solution with the same retention time as desloratadine pyridine nitrogen oxide, the desloratadine pyridine nitrogen oxide content in the test solution can be calculated by peak area using the external standard method.

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