Method for detecting polyethylene glycol in paliperidone palmitate injection

Abstract

The invention belongs to the technical field of medicine quality control, and particularly relates to a method for detecting polyethylene glycol in a paliperidone palmitate injection. The invention relates to a method for detecting polyethylene glycol in a paliperidone palmitate injection. The method comprises the following steps: detecting the content of polyethylene glycol in the paliperidone palmitate injection by using a liquid chromatography method; wherein a detector of the liquid chromatography is a CAD (Computer Aided Design) detector. The detection method disclosed by the invention has the advantages of high sensitivity, strong specificity and accurate detection result.

Description

Technical Field

[0001] The present invention belongs to the technical field of drug quality control, and in particular relates to a method for detecting polyethylene glycol in paliperidone palmitate injection. Background Art

[0002] Paliperidone palmitate injection is a long-acting injection used to treat schizophrenia in adults. The original manufacturer is Janssen Pharmaceuticals Co., Ltd. It has the advantages of sustained release, long-term effect, high safety, and good patient compliance. In the disclosed prescription information of paliperidone palmitate injection, polyethylene glycol 4000 is one of the excipients of its preparation. Other excipients in the prescription include polysorbate 20, citric acid, anhydrous disodium hydrogen phosphate, sodium dihydrogen phosphate monohydrate, and sodium hydroxide. The prescription composition is relatively complex. As a common pharmaceutical polymer excipient, polyethylene glycol 4000 is generally used as a suspending agent or stabilizer to prevent particle aggregation and increase drug stability. Since this excipient will affect the quality of paliperidone palmitate injection, in order to ensure the safety, effectiveness and quality controllability of the preparation, it is necessary to establish a test method for the content of polyethylene glycol 4000 in the preparation.

[0003] When detecting polyethylene glycol 4000, conventional ultraviolet detectors are not applicable because this component has no ultraviolet absorption. The 2020 edition of the Chinese Pharmacopoeia does not include its content determination method. The United States Pharmacopoeia Polyethylene Glycol 3350 and Electrolytes for Oral Solution uses a differential refractometer for determination, and the published literature also includes the use of an evaporative light scattering detector for determination.

[0004] The inventors of the present disclosure found in the research process that for the detection of polyethylene glycol 4000 in paliperidone palmitate injection, the differential refractive index detector is used, the mobile phase preparation is relatively complicated, and reagents such as sodium chloride, potassium chloride, sodium bicarbonate, sodium sulfate, etc. are used, and the sensitivity is not high, which is more suitable for the detection of a single component; the evaporative light scattering detector is used, which also has the problem of low sensitivity, and poor accuracy and repeatability. The prescription ratio of polyethylene glycol 4000 in paliperidone palmitate injection is only 3%, so the above method is not suitable for the detection of polyethylene glycol 4000 in paliperidone palmitate injection.

[0005] Therefore, it is urgent to develop an analytical method with high sensitivity, strong specificity and accurate detection results. Summary of the invention

[0006] The technical problem to be solved by the present disclosure is to develop a method for detecting polyethylene glycol in paliperidone palmitate injection with high sensitivity, strong specificity and accurate detection results, in view of the fact that there is currently no suitable method for detecting polyethylene glycol in paliperidone palmitate injection.

[0007] In order to achieve the above technical objectives, the technical solution adopted by the present disclosure is:

[0008] The present disclosure provides a method for detecting polyethylene glycol in paliperidone palmitate injection, comprising the following steps: using a liquid chromatography method to detect the content of polyethylene glycol in paliperidone palmitate injection; wherein the detector of the liquid chromatography method is a CAD detector. The CAD detector refers to an electrospray detector.

[0009] In some preferred embodiments of the present disclosure, the liquid chromatography method detection step includes: diluting the paliperidone palmitate injection with a diluent to obtain a test solution.

[0010] In some preferred embodiments of the present disclosure, the preparation step of the test solution includes: accurately weighing 50-500 mg of paliperidone palmitate injection, placing it in a 50 ml volumetric bottle, and adding a diluent to dilute to the scale.

[0011] In some preferred embodiments of the present disclosure, the preparation step of the test solution includes: accurately weighing 100-400 mg of paliperidone palmitate injection, placing it in a 50 ml volumetric bottle, and adding a diluent to dilute to the scale.

[0012] In some preferred embodiments of the present disclosure, the preparation step of the test solution includes: accurately weighing 120-300 mg of paliperidone palmitate injection, placing it in a 50 ml volumetric bottle, and adding a diluent to dilute to the scale.

[0013] In some preferred embodiments of the present disclosure, the preparation step of the test solution includes: accurately weighing 150-250 mg of paliperidone palmitate injection, placing it in a 50 ml volumetric bottle, and adding a diluent to dilute to the scale.

[0014] In some preferred embodiments of the present disclosure, the preparation step of the test solution includes: accurately weighing 166-248 mg of paliperidone palmitate injection, placing it in a 50 ml volumetric bottle, and adding a diluent to dilute to the scale.

[0015] In some preferred embodiments of the present disclosure, the preparation step of the test solution includes: accurately weighing 207 mg of paliperidone palmitate injection, placing it in a 50 ml volumetric bottle, and adding a diluent to dilute to the scale.

[0016] In some preferred embodiments of the present disclosure, the liquid chromatography method detection step also includes: detecting the polyethylene glycol in the test solution by liquid chromatography, and calculating the content of polyethylene glycol in the paliperidone palmitate injection according to the concentration-peak area standard curve of polyethylene glycol.

[0017] In some preferred embodiments of the present disclosure, the chromatographic column of the liquid chromatography method is: a reverse phase chromatographic column; preferably, the chromatographic column is a reverse phase C 18 Chromatographic column; preferably, the chromatographic column is a Waters XTerra reverse phase C18 chromatographic column; preferably, the size of the chromatographic column is a Waters XTerra reverse phase C18 chromatographic column 150×3.0mm×3.5μm.

[0018] In some preferred embodiments of the present disclosure, the mobile phase of the liquid chromatography method is: mobile phase A is 0.002-0.02 mol / L ammonium acetate solution; mobile phase B is a mixture of methanol and acetonitrile in a volume ratio of (12-28): (78-88);

[0019] Preferably, the mobile phase A is a 0.005-0.015 mol / L ammonium acetate solution; the mobile phase B is a mixture of methanol and acetonitrile in a volume ratio of (15-25):(75-85);

[0020] Preferably, the mobile phase A is a 0.008-0.012 mol / L ammonium acetate solution; the mobile phase B is a mixture of methanol and acetonitrile in a volume ratio of (18-22):(78-82).

[0021] In some preferred embodiments of the present disclosure, the diluent is a mixed solution of tetrahydrofuran and N,N-dimethylformamide; preferably, the volume ratio of tetrahydrofuran to N,N-dimethylformamide in the diluent is 1-15:85-99; preferably, the volume ratio of tetrahydrofuran to N,N-dimethylformamide in the diluent is 3-8:92-97; preferably, the volume ratio of tetrahydrofuran to N,N-dimethylformamide in the diluent is 4.5-5.5:94.5-95.5; preferably, the volume ratio of tetrahydrofuran to N,N-dimethylformamide in the diluent is 5:95.

[0022] In some preferred embodiments of the present disclosure, the filtering of the CAD detector is 1.0 to 10.0; preferably, the filtering of the CAD detector is 2.0 to 8.0; preferably, the filtering of the CAD detector is 3.0 to 7.0; preferably, the filtering of the CAD detector is 4.0 to 6.0.

[0023] In some preferred embodiments of the present disclosure, the CAD parameters of the CAD detector include: the atomization temperature is Low, the acquisition frequency is 5 to 15 Hz, and the gas partial pressure is 0.1 to 1.0 MPa; preferably, the CAD parameters of the CAD detector include: the atomization temperature is Low, the acquisition frequency is 8 to 12 Hz, and the gas partial pressure is 0.3 to 0.8 MPa; preferably, the CAD parameters of the CAD detector include: the atomization temperature is Low, the acquisition frequency is 10 Hz, and the gas partial pressure is 0.5 MPa.

[0024] In some preferred embodiments of the present disclosure, gradient elution is performed according to the following procedure:

[0025] 0-5min, the proportion of mobile phase A is (80±5)%, and the proportion of mobile phase B is (20±5)%;

[0026] From 5 to 10 min, the proportion of mobile phase A decreased from (80±5)% to (10±5)%, and the proportion of mobile phase B increased from (20±5)% to (90±5)%;

[0027] 10-30 min, the proportion of mobile phase A decreased from (10±5)% to (5±5)%, and the proportion of mobile phase B increased from (90±5)% to (95±5)%;

[0028] From 30 to 30.1 min, the proportion of mobile phase A increased from (5±5)% to (80±5)%, and the proportion of mobile phase B decreased from (95±5)% to (20±5)%;

[0029] 30.1~35min, the proportion of mobile phase A is (80±5)%, and the proportion of mobile phase B is (20±5)%.

[0030] In some preferred embodiments of the present disclosure, gradient elution is performed according to the following procedure:

[0031] 0-5min, the proportion of mobile phase A is (80±2)%, and the proportion of mobile phase B is (20±2)%;

[0032] 5-10 min, the proportion of mobile phase A decreased from (80±2)% to (10±2)%, and the proportion of mobile phase B increased from (20±2)% to (90±2)%;

[0033] 10-30 min, the proportion of mobile phase A decreased from (10±2)% to (5±2)%, and the proportion of mobile phase B increased from (90±2)% to (95±2)%;

[0034] From 30 to 30.1 min, the proportion of mobile phase A increased from (5±2)% to (80±2)%, and the proportion of mobile phase B decreased from (95±2)% to (20±2)%;

[0035] 30.1~35min, the proportion of mobile phase A is (80±2)%, and the proportion of mobile phase B is (20±2)%.

[0036] In some preferred embodiments of the present disclosure, the gradient elution conditions of the liquid chromatography method include: mobile phase A is 0.003-0.018 mol / L ammonium acetate solution; mobile phase B is a mixture of methanol and acetonitrile in a volume ratio of 10-30:70-90; gradient elution is performed according to the following procedure:

[0037] 0-5min, the proportion of mobile phase A is 80%, and the proportion of mobile phase B is 20%;

[0038] 5-10 min, the proportion of mobile phase A decreased from 80% to 10%, and the proportion of mobile phase B increased from 20% to 90%;

[0039] From 10 to 30 min, the proportion of mobile phase A decreased from 10% to 5%, and the proportion of mobile phase B increased from 90% to 95%;

[0040] From 30 to 30.1 min, the proportion of mobile phase A increased from 5% to 80%, and the proportion of mobile phase B decreased from 95% to 20%;

[0041] 30.1~35min, the proportion of mobile phase A is 80%, and the proportion of mobile phase B is 20%.

[0042] In some preferred embodiments of the present disclosure, the gradient elution conditions of the liquid chromatography method include: mobile phase A is 0.005-0.015 mol / L ammonium acetate solution; mobile phase B is a mixture of methanol and acetonitrile in a volume ratio of 15-25:75-85; gradient elution is performed according to the following procedure:

[0043] 0-5min, the proportion of mobile phase A is 80%, and the proportion of mobile phase B is 20%;

[0044] 5-10 min, the proportion of mobile phase A decreased from 80% to 10%, and the proportion of mobile phase B increased from 20% to 90%;

[0045] From 10 to 30 min, the proportion of mobile phase A decreased from 10% to 5%, and the proportion of mobile phase B increased from 90% to 95%;

[0046] From 30 to 30.1 min, the proportion of mobile phase A increased from 5% to 80%, and the proportion of mobile phase B decreased from 95% to 20%;

[0047] 30.1~35min, the proportion of mobile phase A is 80%, and the proportion of mobile phase B is 20%.

[0048] In some preferred embodiments of the present disclosure, the gradient elution conditions of the liquid chromatography method include: mobile phase A is 0.01 mol / L ammonium acetate solution; mobile phase B is a mixture of methanol and acetonitrile in a volume ratio of 20:80; gradient elution is performed according to the following procedure:

[0049] 0-5min, the proportion of mobile phase A is 80%, and the proportion of mobile phase B is 20%;

[0050] 5-10 min, the proportion of mobile phase A decreased from 80% to 10%, and the proportion of mobile phase B increased from 20% to 90%;

[0051] From 10 to 30 min, the proportion of mobile phase A decreased from 10% to 5%, and the proportion of mobile phase B increased from 90% to 95%;

[0052] From 30 to 30.1 min, the proportion of mobile phase A increased from 5% to 80%, and the proportion of mobile phase B decreased from 95% to 20%;

[0053] 30.1~35min, the proportion of mobile phase A is 80%, and the proportion of mobile phase B is 20%.

[0054] In some preferred embodiments of the present disclosure, the gradient elution conditions of the liquid chromatography method include:

[0055]

[0056] Among them, mobile phase A is 0.01 mol / L ammonium acetate solution; mobile phase B is a mixture of methanol and acetonitrile in a volume ratio of 20:80.

[0057] In some preferred embodiments of the present disclosure, the step of preparing a standard curve solution is further included; preferably, the standard curve solution is a gradient concentration solution in the concentration range of 0.09 mg / mL to 0.15 mg / mL; preferably, the step of preparing the standard curve solution includes: taking polyethylene glycol 4000 to prepare a gradient concentration solution of 0.096 mg / mL, 0.108 mg / mL, 0.120 mg / mL, 0.132 mg / mL, and 0.144 mg / mL for establishing a standard curve.

[0058] In some preferred embodiments of the present disclosure, the preparation step of a reference stock solution is further included; preferably, the preparation step of the reference stock solution includes: preparing a reference stock solution of 0.5-3.0 mg / ml; preferably, the preparation step of the reference stock solution includes: preparing a reference stock solution of 0.8-1.5 mg / ml; preferably, the preparation step of the reference stock solution includes: preparing a reference stock solution of 1.1-1.3 mg / ml; preferably, the preparation step of the reference stock solution includes: preparing a reference stock solution of 1.15-1.25 mg / ml.

[0059] In some preferred embodiments of the present disclosure, the column temperature of the liquid chromatography column is 25-45°C; preferably, the column temperature of the liquid chromatography column is 30-40°C; preferably, the column temperature of the liquid chromatography column is 32-38°C; preferably, the column temperature of the liquid chromatography column is 35°C.

[0060] In some preferred embodiments of the present disclosure, the injection volume of the liquid chromatography is 1 to 10 μL; preferably, the injection volume of the liquid chromatography is 2 to 8 μL; preferably, the injection volume of the liquid chromatography is 4 to 6 μL; preferably, the injection volume of the liquid chromatography is 5 μL.

[0061] In some preferred embodiments of the present disclosure, the flow rate of the liquid chromatography is 0.2-1.0 mL / min; preferably, the flow rate of the liquid chromatography is 0.4-0.8 mL / min; preferably, the flow rate of the liquid chromatography is 0.5-0.7 mL / min; preferably, the flow rate of the liquid chromatography is 0.6 mL / min.

[0062] In some preferred embodiments of the present disclosure, the polyethylene glycol is polyethylene glycol 400-8000; preferably, the polyethylene glycol is polyethylene glycol 2000-6000; preferably, the polyethylene glycol is polyethylene glycol 4000.

[0063] The present disclosure has the following advantages:

[0064] (1) Compared with the prior art, the present invention adopts liquid chromatography technology to detect polyethylene glycol in paliperidone palmitate injection, optimizes the chromatographic conditions, has high sensitivity, strong specificity and accurate detection results.

[0065] (2) Compared with the prior art, the present invention adopts liquid chromatography technology to detect polyethylene glycol in paliperidone palmitate injection, optimizes the chromatographic conditions, and has a simple operation method.

[0066] (3) Compared with the prior art, the present invention adopts liquid chromatography technology to detect polyethylene glycol in paliperidone palmitate injection, optimizes the chromatographic conditions, and shortens the analysis time.

[0067] (4) Compared with the prior art, the present invention adopts liquid chromatography technology to detect polyethylene glycol in paliperidone palmitate injection, optimizes the chromatographic conditions, has good reproducibility and strong universality.

[0068] (5) The detection method disclosed in the present invention can be used for the qualitative or quantitative analysis of polyethylene glycol in paliperidone palmitate injection. BRIEF DESCRIPTION OF THE DRAWINGS

[0069] In order to more clearly illustrate the specific embodiments of the present disclosure or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the specific embodiments or the prior art description. Obviously, the drawings described below are some embodiments of the present disclosure. For ordinary technicians in this field, other drawings can also be obtained based on these drawings without creative work. The drawings herein are incorporated into the specification and constitute a part of the specification, showing embodiments that conform to the specification, and are used together with the specification to explain the principles of the specification.

[0070] Figure 1 The liquid chromatogram of Example 2 is shown, wherein the top blue chromatogram elution curve in the figure is the polyethylene glycol 4000 control solution, the middle black chromatogram elution curve is the preparation sample solution, and the bottom pink chromatogram elution curve is the blank solution.

[0071] Figure 2 The liquid chromatogram of Example 3 is shown, wherein the top brown chromatogram elution curve is for the GLIntersil ODS-SP chromatographic column, the middle pink chromatogram elution curve is for the Agilent ZORBAX Eclipse Plus C18 chromatographic column, and the bottom black chromatogram elution curve is for the GL Sciences Intersil ODS-3 chromatographic column.

[0072] Figure 3 The liquid chromatogram under the conditions of chromatographic column 1 and elution gradient 1 in Example 4 is shown.

[0073] Figure 4 The liquid chromatogram under the conditions of chromatographic column 1 and elution gradient 2 in Example 5 is shown.

[0074] Figure 5The liquid chromatogram under the conditions of chromatographic column 2 and elution gradient 2 in Example 6 is shown, wherein the top pink chromatographic elution curve in the figure is the preparation sample solution, the middle blue chromatographic elution curve is the polyethylene glycol 4000 control solution, and the bottom black chromatographic elution curve is the blank solution.

[0075] Figure 6 The liquid chromatogram of Example 7 is shown, wherein the upper blue chromatogram elution curve in the figure is filtering 5.0, and the lower black chromatogram elution curve is filtering 1.0.

[0076] Figure 7 The liquid chromatogram of Comparative Example 1 is shown, wherein the top blue chromatogram elution curve in the figure is a blank solution, the middle green chromatogram elution curve is a polyethylene glycol 4000 control solution, and the bottom purple chromatogram elution curve is a preparation sample solution.

[0077] Figure 8 The liquid chromatogram of Comparative Example 2 is shown, wherein the top green chromatographic elution curve in the figure is the preparation sample solution, the middle blue chromatographic elution curve is the polyethylene glycol 4000 control solution, and the bottom black chromatographic elution curve is the blank solution. DETAILED DESCRIPTION

[0078] Definition and Description

[0079] In order to make it easier to understand the present disclosure, some technical and scientific terms are specifically defined below. In the present disclosure, unless otherwise stated, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. In addition, the cell and tissue culture, microbiology-related terms and laboratory operation steps used herein are terms and routine steps widely used in the corresponding fields. At the same time, in order to better understand the present disclosure, the definition and explanation of the relevant terms are provided below. It should be understood that the present disclosure is not limited to specific methods, reagents, compounds, compositions or biological systems, and of course the above can be changed. It should also be understood that the terms used in the present application are only for describing specific embodiments and are not intended to be limited.

[0080] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise.

[0081] As used herein, the terms "include" and "have" and any variations thereof are intended to cover non-exclusive inclusions. For example, a process, method, device, product or apparatus comprising a series of steps is not limited to the listed steps or modules, but may optionally include steps not listed, or may optionally include other steps inherent to these processes, methods, products or apparatuses.

[0082] In the description of this document, reference is made to “some embodiments”, “some implementation schemes” or “some implementation plans”, which describe a subset of all possible embodiments, but it can be understood that “some embodiments” may be the same subset or different subsets of all possible embodiments and may be combined with each other without conflict.

[0083] CAD detector (electrospray detector) is a universal detector. The detection results of CAD detector are related to the analyte particles, and the signal current is proportional to the mass of the analyte in the sample. Working principle of CAD detector: Step 1: CAD detector converts the analyte into solute particles, and the size of the particles increases with the content of the analyte; Step 2: Solute particles collide with positively charged nitrogen particles, and the charge is transferred to the particles - the larger the solute particles, the more charged they are; Step 3: Solute particles transfer their charge to the collector, and the charge of the solute particles is measured by a highly sensitive electrostatic detector. The resulting signal current is proportional to the content of the solute.

[0084] Example

[0085] In order to make the purpose, technical solution and advantages of the embodiments of the present disclosure clearer, the technical solution in the embodiments of the present disclosure will be clearly and completely described below in conjunction with the drawings in the embodiments of the present disclosure. Obviously, the described embodiments are part of the embodiments of the present disclosure, rather than all the embodiments. The following is only a further description of the present disclosure, and the protection scope of the present disclosure is not limited thereto.

[0086] After searching the pharmacopoeias and related literature of various countries, it was found that the common analytical methods for determining polyethylene glycol 4000 are mostly differential refractive index detectors or evaporative light scattering detectors, both of which have the disadvantage of low sensitivity. Therefore, a new detection method for polyethylene glycol 4000 was developed.

[0087] Embodiment 1:

[0088] 1. Liquid chromatography analysis method

[0089] (1) Instrument parameters

[0090]

[0091]

[0092] (2) Solution preparation:

[0093] Diluent: The volume ratio of tetrahydrofuran to N,N-dimethylformamide is 5:95

[0094] Reference substance stock solution: Take about 60 mg of excipient polyethylene glycol 4000, weigh accurately, put into a 50 ml volumetric flask, dissolve in 5 ml ultrapure water and dilute to the scale with diluent, shake well; can be diluted in equal proportions to make 1.2 mg / ml.

[0095] Preparation of standard curve solution: According to the table below, accurately pipette an appropriate amount of reference stock solution into the corresponding volumetric flask, dilute to the scale with diluent, make up to volume, and shake well.

[0096] Table 1. Preparation of standard curve solutions

[0097]

[0098] Test solution: Take 5 test samples and shake well (shake 30 times within 10 seconds, with a shaking amplitude of about 25 cm and shake for at least 10 seconds), transfer the contents into a stoppered test tube and shake well; accurately weigh about 207 mg of paliperidone palmitate injection, place it in a 50 ml volumetric flask, add diluent and shake to dissolve, dilute to the scale, and shake well.

[0099] (3) Sampling procedure

[0100] Inject samples according to the table below and analyze the test solution after the system suitability requirements are met.

[0101]

[0102] (4) System applicability

[0103] The binary standard curve equation y=ax2+bx+c was drawn based on concentration-peak area, and the correlation coefficient R2≥0.98;

[0104] The program controls the ratio of the standard curve solution L3 to the main peak area in the standard curve solution L3 to be between 95.0% and 105.0%.

[0105] (5) Calculation formula

[0106]

[0107] 2. Methodological validation:

[0108] According to the analysis method determined in Example 1 "1. Liquid chromatography analysis method", the relevant verification test was carried out, and the process and results are as follows:

[0109] (1) Linear (quadratic function)

[0110] According to the analysis method determined in "1. Liquid chromatography analysis method" of Example 1, a control solution was prepared for injection. Due to the particularity of the CAD detector, a quadratic function was used to fit the peak area and concentration.

[0111]

[0112]

[0113] The quadratic function equation of the peak area and polyethylene glycol 4000 concentration is: y=-85.592x2+52.329x-0.1741, R2=0.9999, and the fitting result is good.

[0114] (2) Repeatability

[0115] Repeatable solution: Accurately weigh 207 mg of paliperidone palmitate injection, place in a 50 ml volumetric flask, add diluent and shake to dissolve, dilute to the scale, shake well and prepare 6 copies in parallel.

[0116]

[0117] The RSD% of 6 replicate solutions was 0.9%, no greater than 2%, indicating good repeatability.

[0118] (3) Accuracy

[0119] Prepare according to the table below to contain polyethylene glycol 4000 at a prescription amount of 80%, 90%, 100%, 110%, 120%, 5 levels, and prepare 3 portions in parallel for each level.

[0120]

[0121]

[0122] The measured accuracy results are as follows:

[0123]

[0124] The recovery of polyethylene glycol 4000 was between 100.8% and 104.4%, and the RSD% of the recovery of 15 solutions was 1.2%, indicating good accuracy.

[0125] (4) Solution stability

[0126] Control solution: accurately weigh 60mg polyethylene glycol 4000 into a 50mL volumetric bottle, add 5ml water to dissolve, dilute to scale with diluent and shake well, accurately transfer 5mL to a 50mL volumetric bottle, dilute to scale with diluent and shake well, and prepare two parallel portions. Control solution 1 is placed at room temperature and tested at 0h, 24h, and 48h, respectively. Control solution 2 is placed at 2-8℃ and tested at 0h, 24h, and 48h, respectively.

[0127] Test solution: Take sample 1 and sample 2 solutions under the repeatability item; sample solution 1 is placed at room temperature and tested at 0h, 24h, and 48h, respectively; sample solution 2 is placed at 2-8℃ and tested at 0h, 24h, and 48h, respectively. The results are as follows:

[0128]

[0129] The experimental results show that the reference solution and the test solution have good solution stability at room temperature for 24 hours.

[0130] Embodiment 2:

[0131] The difference from Example 1 is that the instrument parameters are as follows:

[0132]

[0133] The obtained chromatogram is as follows Figure 1 As shown, the top blue chromatographic elution curve in the figure is the polyethylene glycol 4000 control solution, the middle black chromatographic elution curve is the preparation sample solution, and the bottom pink chromatographic elution curve is the blank solution.

[0134] Experimental results: Under the chromatographic conditions, the diluent had no interference with the determination of polyethylene glycol 4000. Polyethylene glycol 4000 appeared in about 6 minutes, and the peak shape and separation from other components were good. However, the theoretical plate number of polyethylene glycol 4000 was only 1876, and the column efficiency was low.

[0135] Embodiment 3:

[0136] The difference from Example 2 is that chromatographic columns of different brands, models, sizes and retention characteristics were used for testing, and other chromatographic conditions remained unchanged. The column efficiency results and chromatograms obtained are as follows:

[0137]

[0138] The obtained chromatogram is as follows Figure 2 As shown, the top brown chromatographic elution curve in the figure is the GL Intersil ODS-SP chromatographic column, the middle pink chromatographic elution curve is the Agilent ZORBAX Eclipse Plus C18 chromatographic column, and the bottom black chromatographic elution curve is the GL Sciences Intersil ODS-3 chromatographic column.

[0139] Experimental results: After replacing different chromatographic columns, the column efficiency has been significantly improved, but it still needs to be further improved.

[0140] Embodiment 4:

[0141] The difference from Example 2 is that the chromatographic parameters are as follows:

[0142]

[0143]

[0144] The obtained chromatogram is as follows Figure 3 shown.

[0145] Experimental results: Under the chromatographic conditions, the retention time of polyethylene glycol 4000 was about 7 minutes, and the running time was shortened to 35 minutes. However, the theoretical plate number of polyethylene glycol 4000 was 1042, and the column efficiency was poor.

[0146] Embodiment 5:

[0147] The difference from Example 2 is that the chromatographic parameters are as follows:

[0148]

[0149] The obtained chromatogram is as follows Figure 4 shown.

[0150] Experimental results: Under the chromatographic conditions, the retention time of polyethylene glycol 4000 was about 10 minutes, the running time was shortened to 35 minutes, and the theoretical plate number reached 24205. The theoretical plate number met expectations, but the baseline was poor, which affected the detection.

[0151] Embodiment 6:

[0152] The difference from Example 2 is that the chromatographic parameters are as follows:

[0153]

[0154]

[0155] The obtained chromatogram is as follows Figure 5 As shown, the top pink chromatographic elution curve in the figure is the preparation sample solution, the middle blue chromatographic elution curve is the polyethylene glycol 4000 control solution, and the bottom black chromatographic elution curve is the blank solution.

[0156] Experimental results: Under this chromatographic condition, the retention time of polyethylene glycol 4000 was about 11 minutes, and the running time was shortened to 35 minutes. The column efficiency was 61032, which met expectations.

[0157] Example 7: CAD detector parameter optimization

[0158] For chromatography Figure 5 After magnification, it was found that the bifurcation at the top of the polyethylene glycol 4000 chromatographic peak was obvious, so the CAD filter of Example 6 was increased from 1.0 to 5.0, and the chromatogram obtained was as follows: Figure 6As shown, the upper blue chromatogram elution curve in the figure is filtering 5.0, and the lower black chromatogram elution curve is filtering 1.0.

[0159] Experimental results: The final chromatographic peak shape of polyethylene glycol 4000 was good and met the detection requirements.

[0160] Example 8: Investigation of the extraction method of the test solution

[0161] Since polyethylene glycol 4000 is dissolved in the solution, the preparation method of using water as a diluent to dissolve the sample (API is insoluble) and then centrifuge to take the supernatant for sampling and testing was investigated. The recovery results are as follows:

[0162]

[0163]

[0164] Results: The recovery rate of polyethylene glycol 4000 decreased with the increase of concentration, indicating that the centrifugation could not completely extract the polyethylene glycol 4000 in the prescription, and a part of it was attached to the surface of the API particles. Therefore, when testing the content of polyethylene glycol 4000, the preparation sample must be completely dissolved before testing. Paliperidone palmitate is easily soluble in tetrahydrofuran, so the volume ratio of tetrahydrofuran to N,N-dimethylformamide (DMF) was 5:95 to dissolve the sample, and the sample was completely dissolved.

[0165] Comparative Example 1: Investigation of the differential detector method

[0166] The difference from Example 2 is that a differential detector is used for analysis and determination, and the chromatographic parameters are as follows:

[0167]

[0168] The obtained chromatogram is as follows Figure 7 As shown, the top blue chromatographic elution curve in the figure is the blank solution, the middle green chromatographic elution curve is the polyethylene glycol 4000 control solution, and the bottom purple chromatographic elution curve is the preparation sample solution.

[0169] Experimental results: Under the chromatographic conditions, neither polyethylene glycol 4000 nor the main component showed any peaks.

[0170] Due to the large difference in polarity between polyethylene glycol 4000 and paliperidone palmitate, the isocratic elution conditions cannot elute the two samples simultaneously. The differential refractometer detector is suitable for isocratic methods but not for gradient methods. Therefore, the detector is not applicable to this project.

[0171] Comparative Example 2: ELSD detector method investigation

[0172] The difference from Example 2 is that the ELSD detector is used for analysis and determination, and the chromatographic parameters are as follows:

[0173]

[0174]

[0175] The obtained chromatogram is as follows Figure 8 As shown, the top green chromatographic elution curve in the figure is the preparation sample solution, the middle blue chromatographic elution curve is the polyethylene glycol 4000 control solution, and the bottom black chromatographic elution curve is the blank solution.

[0176] Experimental results: Under this chromatographic condition, polyethylene glycol 4000 peaked at about 7 minutes, but the response was very weak and could not be integrated. The method had poor sensitivity and was not suitable for the determination of polyethylene glycol 4000 in paliperidone palmitate injection.

[0177] The foregoing description of specific exemplary embodiments of the present disclosure is for the purpose of illustration and demonstration. These descriptions are not intended to limit the present disclosure to the precise form disclosed, and it is clear that many changes and variations can be made based on the above teachings. The purpose of selecting and describing the exemplary embodiments is to explain the specific principles of the present disclosure and its practical application, so that those skilled in the art can realize and utilize various different exemplary embodiments of the present disclosure and various different selections and changes. The scope of the present disclosure is intended to be defined by the claims and their equivalents.

Claims

1. A method for detecting polyethylene glycol in paliperidone palmitate injection, comprising the following steps: The content of polyethylene glycol in paliperidone palmitate injection is detected by liquid chromatography; wherein the detector of the liquid chromatography is a CAD detector.

2. The detection method according to claim 1, characterized in that: The liquid chromatography method detection step comprises: diluting the paliperidone palmitate injection with a diluent to obtain a test solution; Preferably, the diluent is a mixed solution of tetrahydrofuran and N,N-dimethylformamide; Preferably, the volume ratio of tetrahydrofuran to N,N-dimethylformamide in the diluent is (1-15):(85-99); Preferably, the volume ratio of tetrahydrofuran to N,N-dimethylformamide in the diluent is (3-8):(92-97); preferably, the volume ratio of tetrahydrofuran to N,N-dimethylformamide in the diluent is (4.5-5.5):(94.5-95.5); preferably, the volume ratio of tetrahydrofuran to N,N-dimethylformamide in the diluent is 5:

95.

3. The detection method according to claim 1 or 2, characterized in that: The chromatographic column of the liquid chromatography method is: a reverse phase chromatographic column; Preferably, the chromatographic column is a reverse phase C 18 Chromatographic columns; Preferably, the chromatographic column is a Waters XTerra reverse phase C18 chromatographic column; Preferably, the size of the chromatographic column is a Waters XTerra reverse phase C18 chromatographic column 150×3.0 mm×3.5 μm.

4. The detection method according to any one of claims 1 to 3, characterized in that: The filtering of the CAD detector is 1.0 to 10.0; preferably, the filtering of the CAD detector is 2.0 to 8.0; preferably, the filtering of the CAD detector is 3.0 to 7.0; preferably, the filtering of the CAD detector is 4.0 to 6.

0.

5. The detection method according to any one of claims 1 to 4, characterized in that: The liquid chromatography detection step also includes: detecting the polyethylene glycol in the test solution by liquid chromatography, and calculating the content of polyethylene glycol in the paliperidone palmitate injection according to the concentration-peak area standard curve of polyethylene glycol.

6. The detection method according to any one of claims 1 to 5, characterized in that: The mobile phase of the liquid chromatography method is: mobile phase A is 0.002-0.02 mol / L ammonium acetate solution; mobile phase B is a mixture of methanol and acetonitrile in a volume ratio of (12-28): (78-88); Preferably, the mobile phase A is a 0.005-0.015 mol / L ammonium acetate solution; the mobile phase B is a mixture of methanol and acetonitrile in a volume ratio of (15-25):(75-85); Preferably, the mobile phase A is a 0.008-0.012 mol / L ammonium acetate solution; the mobile phase B is a mixture of methanol and acetonitrile in a volume ratio of (18-22):(78-82).

7. The detection method according to any one of claims 2 to 6, characterized in that: in, The preparation steps of the test solution include: accurately weighing 150-250 mg of paliperidone palmitate injection, placing it in a 50 ml volumetric bottle, and adding a diluent to dilute to the scale; Preferably, the preparation step of the test solution comprises: accurately weighing 166-248 mg of paliperidone palmitate injection, placing it in a 50 ml volumetric bottle, and adding a diluent to dilute to the scale; Preferably, the preparation step of the test solution comprises: accurately weighing 207 mg of paliperidone palmitate injection, placing it in a 50 ml volumetric bottle, and adding a diluent to dilute to the scale.

8. The detection method according to any one of claims 1 to 7, characterized in that: The CAD parameters of the CAD detector include: atomization temperature is Low, acquisition frequency is 5-15Hz, and gas partial pressure is 0.1-1.0MPa; preferably, the CAD parameters of the CAD detector include: atomization temperature is Low, acquisition frequency is 8-12Hz, and gas partial pressure is 0.3-0.8MPa; preferably, the CAD parameters of the CAD detector include: atomization temperature is Low, acquisition frequency is 10Hz, and gas partial pressure is 0.5MPa.

9. The detection method according to any one of claims 1 to 8, characterized in that: The gradient elution conditions of the liquid chromatography method include: mobile phase A is 0.003-0.018 mol / L ammonium acetate solution; mobile phase B is a mixture of methanol and acetonitrile in a volume ratio of 10-30:70-90; gradient elution is performed according to the following procedure: 0-5min, the proportion of mobile phase A is 80%, and the proportion of mobile phase B is 20%; 5-10 min, the proportion of mobile phase A decreased from 80% to 10%, and the proportion of mobile phase B increased from 20% to 90%; From 10 to 30 min, the proportion of mobile phase A decreased from 10% to 5%, and the proportion of mobile phase B increased from 90% to 95%; From 30 to 30.1 min, the proportion of mobile phase A increased from 5% to 80%, and the proportion of mobile phase B decreased from 95% to 20%; 30.1-35min, the proportion of mobile phase A is 80%, and the proportion of mobile phase B is 20%; Preferably, the gradient elution conditions of the liquid chromatography method include: mobile phase A is 0.005-0.015 mol / L ammonium acetate solution; mobile phase B is a mixture of methanol and acetonitrile in a volume ratio of 15-25:75-85; gradient elution is performed according to the following procedure: 0-5min, the proportion of mobile phase A is 80%, and the proportion of mobile phase B is 20%; 5-10 min, the proportion of mobile phase A decreased from 80% to 10%, and the proportion of mobile phase B increased from 20% to 90%; From 10 to 30 min, the proportion of mobile phase A decreased from 10% to 5%, and the proportion of mobile phase B increased from 90% to 95%; From 30 to 30.1 min, the proportion of mobile phase A increased from 5% to 80%, and the proportion of mobile phase B decreased from 95% to 20%; 30.1-35min, the proportion of mobile phase A is 80%, and the proportion of mobile phase B is 20%; Preferably, the gradient elution conditions of the liquid chromatography method include: mobile phase A is 0.01 mol / L ammonium acetate solution; mobile phase B is a mixture of methanol and acetonitrile in a volume ratio of 20:80; gradient elution is performed according to the following procedure: 0-5min, the proportion of mobile phase A is 80%, and the proportion of mobile phase B is 20%; 5-10 min, the proportion of mobile phase A decreased from 80% to 10%, and the proportion of mobile phase B increased from 20% to 90%; From 10 to 30 min, the proportion of mobile phase A decreased from 10% to 5%, and the proportion of mobile phase B increased from 90% to 95%; From 30 to 30.1 min, the proportion of mobile phase A increased from 5% to 80%, and the proportion of mobile phase B decreased from 95% to 20%; 30.1~35min, the proportion of mobile phase A is 80%, and the proportion of mobile phase B is 20%.

10. The detection method according to any one of claims 1 to 9, characterized in that: in, The method further comprises the step of preparing a standard curve solution; preferably, the standard curve solution is a gradient concentration solution in the concentration range of 0.09 mg / mL to 0.15 mg / mL; preferably, the step of preparing the standard curve solution comprises: taking polyethylene glycol 4000 to prepare a gradient concentration solution of 0.096 mg / mL, 0.108 mg / mL, 0.120 mg / mL, 0.132 mg / mL, and 0.144 mg / mL for establishing a standard curve; Preferably, the method further comprises the step of preparing a reference stock solution; preferably, the step of preparing the reference stock solution comprises: preparing a reference stock solution of 0.5 to 3.0 mg / ml; preferably, the step of preparing the reference stock solution comprises: preparing a reference stock solution of 0.8 to 1.5 mg / ml; preferably, the step of preparing the reference stock solution comprises: preparing a reference stock solution of 1.1 to 1.3 mg / ml; preferably, the step of preparing the reference stock solution comprises: preparing a reference stock solution of 1.15 to 1.25 mg / ml; Preferably, the column temperature of the chromatographic column of the liquid chromatography is 25-45°C; preferably, the column temperature of the chromatographic column of the liquid chromatography is 30-40°C; preferably, the column temperature of the chromatographic column of the liquid chromatography is 32-38°C; preferably, the column temperature of the chromatographic column of the liquid chromatography is 35°C; Preferably, the injection volume of the liquid chromatography is 1 to 10 μL; preferably, the injection volume of the liquid chromatography is 2 to 8 μL; preferably, the injection volume of the liquid chromatography is 4 to 6 μL; preferably, the injection volume of the liquid chromatography is 5 μL; Preferably, the flow rate of the liquid chromatography is 0.2 to 1.0 mL / min; preferably, the flow rate of the liquid chromatography is 0.4 to 0.8 mL / min; preferably, the flow rate of the liquid chromatography is 0.5 to 0.7 mL / min; preferably, the flow rate of the liquid chromatography is 0.6 mL / min; Preferably, the polyethylene glycol is polyethylene glycol 400-8000; preferably, the polyethylene glycol is polyethylene glycol 2000-6000; preferably, the polyethylene glycol is polyethylene glycol 4000.

Images