High performance liquid chromatography method for efficiently separating and detecting isomeride impurities in fluvoxamine maleate

Abstract

A high performance liquid chromatography method for efficiently separating and detecting a class of isomeride impurities in fluvoxamine maleate and / or a derivative thereof and a preparation thereof adopts octyl silane bonded silica gel as a filler, the column temperature is 20-35 DEG C, the detection wavelength is 240-260 nm, a mobile phase consists of a phosphate solution and an organic solvent, the pH value of the mobile phase is 3.0-4.0, the flow velocity of the mobile phase is 0.8-1.2 m1 / min, the column temperature is 20-35 DEG C, the detection wavelength is 240-260 nm, and the detection time is 20-30 minutes. The temperature of the sample chamber is 5 DEG C; wherein the isomeride impurities are a compound shown in a formula 1 and a compound shown in a formula 2. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the technical field of medicines, and in particular relates to a high performance liquid chromatography method for efficiently separating and detecting isomeric impurities in fluvoxamine maleate. Background Art

[0002] Fluvoxamine Maleate ((E)-5-methoxy-4'-trifluoromethylvaleriophenone-oxy-(2-aminoacetyl oxime) maleate) is a 5-hydroxytryptamine reuptake inhibitor that acts on brain nerve cells. It is clinically used to treat depression and related symptoms and obsessive-compulsive disorder symptoms. Its advantages are that it has no excitatory or sedative effects, no anticholinergic or antihistamine effects, and does not affect the activity of monoamine oxidase (MAO). It has no effect on the cardiovascular system and does not cause orthostatic hypotension.

[0003] Fluvoxamine maleate structure

[0004]

[0005] Effective control of key impurities in drugs is directly related to the quality, safety and effectiveness of drugs. In impurity detection, isomeric impurities are often difficult to separate and detect due to their similar structures and polarities. Finding a chromatographic condition that can effectively separate and detect isomeric impurities is conducive to achieving controllable drug quality and ensuring the effectiveness and safety of drugs. Summary of the invention

[0006] The purpose of the present invention is to provide a high-performance liquid chromatography method for efficiently separating and detecting isomeric impurities in fluvoxamine maleate and / or its derivatives and preparations thereof, wherein octylsilane bonded silica gel is used as a filler, the column temperature is 20-35° C., the detection wavelength is 240-260 nm, the mobile phase is composed of a phosphate solution and an organic solvent, the pH of the mobile phase is 3.0-4.0, the flow rate of the mobile phase is 0.8-1.2 ml / min, and the sample chamber temperature is 5° C. Wherein, the isomeric impurities are compounds of formula 1 and compounds of formula 2.

[0007]

[0008] In a preferred technical solution of the present invention, the detector is selected from any one of a diode array detector or an ultraviolet detector.

[0009] In a preferred technical solution of the present invention, the column temperature is selected from any one of 20°C, 22°C, 25°C, 28°C, 30°C, and 35°C, preferably 35°C.

[0010] Preferably, the sample chamber temperature is 5°C.

[0011] In the preferred technical solution of the present invention, the flow rate of the mobile phase is selected from any one of 0.8 ml / min, 0.9 ml / min, 1.0 ml / min, 1.1 ml / min, and 1.2 ml / min.

[0012] In a preferred technical solution of the present invention, the detection wavelength is selected from any one of 240nm, 243nm, 248nm, 254nm, and 260nm.

[0013] In the preferred technical solution of the present invention, the salt solution constituting the mobile phase is a phosphate solution, and the phosphate is preferably any one or a combination of sodium phosphate, potassium phosphate, and ammonium phosphate.

[0014] In a preferred technical solution of the present invention, the phosphate is selected from any one of dipotassium hydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, sodium monohydrogen phosphate, ammonium monohydrogen phosphate, diammonium hydrogen phosphate, sodium octane sulfonate, sodium heptane sulfonate, and sodium hexane sulfonate, or a combination thereof.

[0015] In a preferred technical solution of the present invention, the organic solvent constituting the mobile phase is selected from any one of acetonitrile and methanol or a combination thereof.

[0016] In a preferred technical solution of the present invention, the pH of the mobile phase is selected from any one of 3.0, 3.5, and 4.0.

[0017] In a preferred technical solution of the present invention, the pH regulator is selected from any one or a combination of hydrochloric acid, phosphoric acid, sodium hydroxide and ammonia water.

[0018] In a preferred technical solution of the present invention, the dilution solvent is selected from one or more of water, methanol, acetonitrile, acetonitrile aqueous solution, and mobile phase.

[0019] In the preferred technical solution of the present invention, the diluting solvent is used to configure, dissolve, and dilute any one of the test solution, the control solution, and the reference substance solution.

[0020] In a preferred technical solution of the present invention, the diameter of the chromatographic column is 3-5 mm, the length of the chromatographic column is 100-300 mm, and the particle size of the filler in the chromatographic column is 3-5 μm.

[0021] In a preferred technical solution of the present invention, the chromatographic column is selected from Kromasil 100-5-C8, 4.6×250 mm, 5 μm.

[0022] In the preferred technical scheme of the present invention, octylsilane bonded silica gel is selected as the reverse phase column, and phosphate buffer (take 5.0g of diammonium hydrogen phosphate and 1.0g of sodium heptane sulfonate, add 400ml of water to dissolve)-methanol (46:54), adjusted to pH 3.5 with phosphoric acid as the mobile phase, the column temperature is 35°C, the detection wavelength is 254nm, the flow rate of the mobile phase is 1.0m1 / min, the injection volume is 20μl, and the sample chamber temperature is 5°C.

[0023] In a preferred technical solution of the present invention, the derivative of fluvoxamine maleate is selected from any one of the pharmaceutically acceptable salts, isomers and solvates of fluvoxamine maleate.

[0024] In the preferred technical scheme of the present invention, the preparation of fluvoxamine maleate is selected from the preparations prepared by conventional methods in the art, preferably selected from tablets, dispersible tablets, enteric-coated tablets, lozenges, chewable tablets, effervescent tablets, orally disintegrating tablets, sustained-release tablets, controlled-release tablets, capsules, granules, powders, oral liquids, injections, lyophilized powder injections, sprays, pills, micropills, syrups, suspensions, effervescent agents, injections, and lyophilized powder injections.

[0025] The compound of formula 1 of the present invention is the reaction product of fluvoxamine and maleic acid.

[0026] The reaction equation is as follows

[0027]

[0028] In the detection of related substances in fluvoxamine maleate tablets, it was found that under extremely hot conditions (high temperature 105°C, 2h), the content of the compound of formula 1 (relative retention time 1.53) increased significantly.

[0029] The compound of formula 2 of the present invention is the reaction product of fluvoxamine and sodium stearyl fumarate.

[0030] The reaction equation is as follows

[0031]

[0032] The present invention provides a high performance liquid chromatography method for efficiently separating and detecting isomeric impurities in fluvoxamine maleate or its derivatives and preparations thereof. The method scientifically screens the chromatographic conditions and elution conditions of the high performance liquid chromatography method, realizes the efficient separation and detection of the main component and isomeric impurities, is conducive to better realizing the quality control of drugs, and ensures the effectiveness and safety of drugs.

[0033] The method of the invention has the advantages of being simple to operate, stable and reliable, and suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Attached Figure 1is the HPLC-MS / MS of the compound of formula 1;

[0035] Attached Figure 2 Liquid chromatogram for efficient separation and detection of isomeric impurities in fluvoxamine maleate tablets. DETAILED DESCRIPTION

[0036] The above contents of the present invention are further described in detail below through specific implementation methods, but this should not be understood as any limitation to the subject matter of the present invention. All technical solutions implemented based on the above contents of the present invention belong to the scope of the present invention.

[0037] Example 1 Methodological Validation - Specificity

[0038] (1) Chromatographic conditions

[0039] Detector: UV detector

[0040] Chromatographic column: Octylsilane bonded silica gel (Kromasil 100-5-C8, 4.6×250mm, 5μm)

[0041] Mobile phase: phosphate buffer (take 5.0 g of diammonium hydrogen phosphate and 1.0 g of sodium heptane sulfonate, add 400 ml of water to dissolve)-methanol (46:54), adjust the pH to 3.5 with phosphoric acid.

[0042] Column temperature: 35°C

[0043] Detection wavelength: 254nm

[0044] Flow rate: 1.0m1 / min

[0045] Injection volume 20 μl

[0046] Sample chamber temperature: 5°C

[0047] Blank solution: diluent

[0048] Diluent: Mobile phase

[0049] Calculation formula:

[0050]

[0051] Where:

[0052] A pair: the peak area of ​​the main component in the control solution;

[0053] A impurities: impurity peak area in the test solution;

[0054] f: Correction factor for each impurity

[0055] 100: Dilution factor of control solution, ml.

[0056] Experimental procedures

[0057] Blank solution: diluent

[0058] Stock solution of the reference substance of the compound of formula 1: Accurately weigh 5 mg of the reference substance solution of the impurity compound of formula 1, accurately weigh, place in a 25 ml volumetric flask, dissolve and dilute to the scale with the mobile phase, shake well, and obtain.

[0059] Stock solution of reference substance of compound of formula 2: Accurately weigh 5 mg of reference substance solution of compound of formula 2, accurately weigh, place in a 25 ml volumetric flask, dissolve and dilute to scale with mobile phase, shake well, and obtain

[0060] Blank excipient solution: Take an appropriate amount of blank excipient of fluvoxamine maleate tablets (approximately equivalent to 50 mg of fluvoxamine maleate), place it in a 50 ml volumetric flask, dissolve and dilute to the scale with mobile phase, shake well, filter, and obtain.

[0061] Formula 1 compound positioning solution: Take an appropriate amount of Formula 1 compound reference substance stock solution, dilute it with mobile phase to a suitable concentration, shake well, and obtain.

[0062] Formula 2 compound positioning solution: Take an appropriate amount of Formula 2 compound reference substance stock solution, dilute it with mobile phase to a suitable concentration, shake well, and obtain.

[0063] Test solution: Take an appropriate amount of fine powder of this product (approximately equivalent to 50 mg of fluvoxamine maleate), accurately weigh, place in a 50 ml volumetric flask, add an appropriate amount of mobile phase, ultrasonicate for 10 minutes, dilute to the scale with mobile phase, shake well, filter, and obtain.

[0064] Spiked test solution: Take an appropriate amount of fine powder of this product (approximately equivalent to 50 mg of fluvoxamine maleate), accurately weigh, place in a 50 ml volumetric flask, add an appropriate amount of mobile phase, sonicate for 10 minutes, accurately add an appropriate amount of reference stock solution of the compound of formula 1 and the reference stock solution of the compound of formula 2 into the above volumetric flask, dilute to the scale with mobile phase, shake well, filter, and obtain.

[0065] Take appropriate amount of blank solution, impurity location solution, blank excipient solution, test solution, and spiked test solution and inject according to chromatographic conditions.

[0066] Result: No interference in blank; good separation of impurities.

[0067] Example 2 Methodological Validation - Limit of Quantitation

[0068] Chromatographic conditions: Same as Example 1

[0069] Stock solution of the reference substance of the compound of formula 1: Accurately weigh 5 mg of the reference substance solution of the impurity compound of formula 1, accurately weigh, place in a 25 ml volumetric flask, dissolve and dilute to the scale with the mobile phase, shake well, and obtain.

[0070] Stock solution of reference substance of compound of formula 2: Accurately weigh 5 mg of reference substance solution of compound of formula 2, accurately weigh, place in a 25 ml volumetric flask, dissolve and dilute to scale with mobile phase, shake well, and obtain

[0071] Quantitative limit solution: Take appropriate amount of reference substance stock solution of compound of formula 1 and reference substance stock solution of compound of formula 2, dilute to appropriate concentration with mobile phase, shake well, and obtain.

[0072] Requirements: S / N should be between 10 and 20.

[0073] Take blank solution and appropriate amount of quantitative limit solution and inject according to chromatographic conditions.

[0074] Results: The quantitative limit concentration of the compound of formula 1 was 0.2166 μg / ml, S / N=17.0; the quantitative limit concentration of the compound of formula 2 was 0.2036 μg / ml, S / N=14.3.

[0075] Example 3 Methodological Validation - Detection Limit

[0076] Chromatographic conditions: Same as Example 1

[0077] Stock solution of the reference substance of the compound of formula 1: Accurately weigh 5 mg of the reference substance solution of the impurity compound of formula 1, accurately weigh, place in a 25 ml volumetric flask, dissolve and dilute to the scale with the mobile phase, shake well, and obtain.

[0078] Stock solution of reference substance of compound of formula 2: Accurately weigh 5 mg of reference substance solution of compound of formula 2, accurately weigh, place in a 25 ml volumetric flask, dissolve and dilute to scale with mobile phase, shake well, and obtain

[0079] Detection limit solution: Take an appropriate amount of quantitation limit solution, dilute it with mobile phase to an appropriate concentration, shake well, and you have it.

[0080] Requirements: S / N should be between 3 and 10.

[0081] Take appropriate amount of blank solution and detection limit solution and inject according to chromatographic conditions.

[0082] Results: The detection limit concentration of the compound of formula 1 was 0.0650 μg / ml, S / N=8.6; the detection limit concentration of the compound of formula 2 was 0.0641 μg / ml, S / N=7.3.

[0083] Example 4 Methodological Validation - Repeatability

[0084] Chromatographic conditions: Same as Example 1

[0085] Stock solution of the reference substance of the compound of formula 1: Accurately weigh 5 mg of the reference substance solution of the impurity compound of formula 1, accurately weigh, place in a 25 ml volumetric flask, dissolve and dilute to the scale with the mobile phase, shake well, and obtain.

[0086] Stock solution of reference substance of compound of formula 2: Accurately weigh 5 mg of reference substance solution of compound of formula 2, accurately weigh, place in a 25 ml volumetric flask, dissolve and dilute to scale with mobile phase, shake well, and obtain

[0087] Repeatability solution: Take an appropriate amount of fine powder of this product (equivalent to 50 mg of fluvoxamine maleate), weigh accurately, place in a 50 ml volumetric bottle, add an appropriate amount of mobile phase, sonicate for 10 minutes, accurately add an appropriate amount of reference substance stock solution of the compound of formula 1 and the reference substance stock solution of the compound of formula 2 to the above volumetric bottle, dilute to the scale with mobile phase, shake well, filter, and obtain. (Prepare 6 portions in parallel)

[0088] Control solution: Take an appropriate amount of the test solution and dilute it 100 times.

[0089] Take appropriate amount of blank solution, reproducible solution and control solution and inject according to chromatographic conditions.

[0090] Results: The RSD of the impurity content of the compound of formula 1 in the 6-shot repeatability sample solution was 2.6%, and the RSD of the impurity content of the compound of formula 1 was 4.3%, with good repeatability.

[0091] Example 5 Methodological Validation - Accuracy

[0092] Chromatographic conditions: Same as Example 1

[0093] Stock solution of the reference substance of the compound of formula 1: Accurately weigh 5 mg of the reference substance solution of the impurity compound of formula 1, accurately weigh, place in a 25 ml volumetric flask, dissolve and dilute to the scale with the mobile phase, shake well, and obtain.

[0094] Stock solution of reference substance of compound of formula 2: Accurately weigh 5 mg of reference substance solution of compound of formula 2, accurately weigh, place in a 25 ml volumetric flask, dissolve and dilute to scale with mobile phase, shake well, and obtain

[0095] Accuracy solution -80%: Take an appropriate amount of fine powder of this product (equivalent to 50mg of fluvoxamine maleate), weigh accurately, place in a 50ml volumetric bottle, add an appropriate amount of mobile phase, sonicate for 10 minutes, accurately add an appropriate amount of reference stock solution of compound 1 and reference stock solution of compound 2 to the above volumetric bottle, dilute to the scale with mobile phase, shake well, filter, and obtain. (Prepare 3 copies in parallel)

[0096] Accuracy solution - 100%: Take an appropriate amount of fine powder of this product (equivalent to 50 mg of fluvoxamine maleate), weigh accurately, place in a 50 ml volumetric bottle, add an appropriate amount of mobile phase, sonicate for 10 minutes, accurately add an appropriate amount of reference substance stock solution of compound of formula 1 and reference substance stock solution of compound of formula 2 to the above volumetric bottle, dilute to the scale with mobile phase, shake well, filter, and obtain. (Prepare 3 portions in parallel)

[0097] Accuracy solution -120%: Take an appropriate amount of fine powder of this product (equivalent to 50mg of fluvoxamine maleate), weigh accurately, place in a 50ml volumetric bottle, add an appropriate amount of mobile phase, sonicate for 10 minutes, accurately add an appropriate amount of reference substance stock solution of compound of formula 1 and reference substance stock solution of compound of formula 2 to the above volumetric bottle, dilute to the scale with mobile phase, shake well, filter, and obtain. (Prepare 3 portions in parallel)

[0098] Control solution: Take an appropriate amount of the test solution and dilute it 100 times.

[0099] Take appropriate amount of blank solution, accuracy solution and reference solution and inject according to chromatographic conditions.

[0100] Results: In 9 accuracy solutions, the average recovery of the compound of formula 1 was 92%, RSD = 3.9, and the average recovery of the compound of formula 1 was 94%, RSD = 2.7, with good accuracy.

[0101] Example 6 Methodological Verification - Solution Stability

[0102] Chromatographic conditions: Same as Example 1

[0103] Spiked test solution: same as Example 1

[0104] Take an appropriate amount of the spiked test solution and inject it at different time points according to the chromatographic conditions to examine the stability of the solution.

[0105] Results: The solution had good stability within 48 hours.

[0106] The above embodiments are only used to help understand the method and core idea of ​​the present invention. It should be noted that, for those skilled in the art, several improvements and modifications can be made to the present invention without departing from the principles of the present invention, and these improvements and modifications also fall within the scope of protection of the claims of the present invention.

Claims

1. A high performance liquid chromatography method for efficiently separating and detecting isomeric impurities in fluvoxamine maleate and / or its derivatives and preparations thereof, characterized in that: Octylsilane bonded silica gel is used as a filler, the column temperature is 20-35°C, the detection wavelength is 240-260nm, the mobile phase is composed of a phosphate solution and an organic solvent, the pH of the mobile phase is 3.0-4.0, the flow rate of the mobile phase is 0.8-1.2ml / min, and the sample chamber temperature is 5°C. Wherein, the isomeric impurities are compounds of formula 1 and compounds of formula 2.

2. The high performance liquid chromatography method according to claim 1, characterized in that The column temperature is selected from any one of 20°C, 22°C, 25°C, 28°C, 30°C, and 35°C.

3. The high performance liquid chromatography method according to claim 1, characterized in that The flow rate of the mobile phase was any one of 0.8 ml / min, 0.9 ml / min, 1.0 ml / min, 1.1 ml / min, and 1.2 ml / min.

4. The high performance liquid chromatography method according to claim 1, characterized in that The detection wavelength is selected from any one of 240 nm, 243 nm, 248 nm, 254 nm, and 260 nm.

5. The high performance liquid chromatography method according to claim 1, characterized in that The salt solution constituting the mobile phase is a phosphate solution, and the phosphate is preferably any one or a combination of sodium phosphate, potassium phosphate, and ammonium phosphate.

6. The high performance liquid chromatography method according to claim 1, characterized in that The phosphate is selected from any one of dipotassium hydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, sodium monohydrogen phosphate, ammonium monohydrogen phosphate, diammonium hydrogen phosphate, sodium octane sulfonate, sodium heptane sulfonate, sodium hexane sulfonate or a combination thereof.

7. The high performance liquid chromatography method according to claim 1, characterized in that The organic solvent constituting the mobile phase is selected from any one of acetonitrile and methanol or a combination thereof.

8. The high performance liquid chromatography method according to claim 1, characterized in that The mobile phase pH is selected from any one of 3.0, 3.5, and 4.

0.

9. The high performance liquid chromatography method according to claim 1, characterized in that The derivative of fluvoxamine maleate is selected from any one of the pharmaceutically acceptable salts, isomers and solvates of fluvoxamine maleate.

10. The high performance liquid chromatography method according to claim 1, characterized in that The preparation of fluvoxamine maleate is selected from preparations prepared by conventional methods in the art, preferably selected from tablets, dispersible tablets, enteric-coated tablets, lozenges, chewable tablets, effervescent tablets, orally disintegrating tablets, sustained-release tablets, controlled-release tablets, capsules, granules, powders, oral liquids, injections, lyophilized powder injections, sprays, pills, micropills, syrups, suspensions, effervescent agents, injections, and lyophilized powder injections.

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