Method for testing oral amisulpride solution

Abstract

The invention relates to the technical field of detection, and discloses an amisulpride oral solution detection method, which comprises: S1, sample pretreatment: sequentially carrying out activation, sample loading, elution and elution operation on an oral solution through a solid phase extraction column, and collecting the eluate as a purification liquid; s2, chromatography and mass spectrometry detection: injecting the purified liquid obtained in S1 into an ultra-high performance liquid chromatography and triple quadrupole mass spectrometry combined system; s3, preparing a microorganism test solution, namely mixing the oral solution and the neutralizer solution according to a volume ratio; s4, microbial limit inspection; and S5, preservative detection. The same test solution is synchronously used for microorganism and chemical detection, two key analyses are completed through one-time preparation, compared with a mode that two samples need to be treated separately in a traditional method, the problem of system error accumulation caused by operation redundancy is thoroughly solved, and the scheme remarkably improves the detection efficiency on the premise of ensuring the data accuracy.

Description

Technical Field

[0001] The invention relates to the field of detection technology, in particular to a method for detecting an amisulpride oral solution. Background Art

[0002] Amisulpride is a core drug for the treatment of schizophrenia. Its oral solution dosage form is increasingly widely used in clinical applications because it is convenient for dose adjustment and use by patients with dysphagia. The quality control of this type of preparation must simultaneously meet multiple indicator detection requirements such as the main drug content, specific impurities, microbial limits and preservative content. Existing inspection methods usually refer to the general principles of the pharmacopoeia and use independent processes to process chemical and microbial samples separately, involving technical modules such as solid phase extraction purification, chromatography-mass spectrometry analysis, and membrane filtration colony culture.

[0003] The current technical solution requires that microbial inspection and preservative testing must use two physically isolated test fluid samples. This separation processing mode results in repeated preparation processes causing sample consumption to double, significantly increasing the risk of wasting precious clinical samples and seriously restricting the overall reliability of the test results. Summary of the Invention

[0004] In response to the shortcomings of the existing technology, the present invention provides a method for testing amisulpride oral solution, which solves the problem that repeated preparation processes cause sample consumption to double, significantly increases the risk of wasting precious clinical samples, and seriously restricts the overall reliability of test results.

[0005] To achieve the above object, the present invention is implemented by the following technical solution: a method for testing amisulpride oral solution, comprising the following steps:

[0006] S1. Sample pretreatment: The oral solution was activated, loaded, washed, and eluted through a solid phase extraction column in sequence, and the eluate was collected as the purification solution;

[0007] S2. Chromatography and mass spectrometry detection: The purified solution obtained in S1 is injected into an ultra-high performance liquid chromatography coupled with a triple quadrupole mass spectrometry system to detect the content of amisulpride and specific impurities under the set chromatographic and mass spectrometry conditions;

[0008] S3. Preparation of microbial test solution: mixing the oral solution with a neutralizer solution according to a volume ratio, wherein the neutralizer comprises histidine and sodium thiosulfate;

[0009] S4. Microbial limit test: Take the test solution obtained in S3 and perform membrane filtration, washing and colony culture;

[0010] S5. Preservative detection: Take the test solution obtained in S3 and test the sodium benzoate content.

[0011] Preferably, in S1, activation, sample loading, washing and elution include the following specific steps:

[0012] The activation operation uses 3-7 mL of methanol and 3-7 mL of pure water;

[0013] The elution operation uses 3-7 mL of pure water and 3-7 mL of an aqueous solution containing 3-7% methanol in sequence;

[0014] The elution operation uses a mixed solvent of 3-7 mL of methanol and acetonitrile, with a volume ratio of 1:0.8-1.2:1.4.

[0015] Preferably, in S1, the eluate is concentrated and then diluted to 1.5-2.5 mL with 8-12 mM ammonium acetate solution containing 0.05-0.2% formic acid.

[0016] Preferably, in said S2, the chromatographic conditions include:

[0017] Chromatographic column: C18 column with an inner diameter of 1.8-2.3 mm and a length of 80-120 mm, and a filler particle size of 1.5-2.0 μm;

[0018] Mobile phase A: 8-12 mM ammonium acetate solution containing 0.05-0.2% formic acid;

[0019] Mobile phase B contained 0.05-0.2% formic acid in acetonitrile.

[0020] Preferably, in said S2, the mass spectrometry conditions set include:

[0021] Monitoring ion pair parameters:

[0022] Amisulpride parent ion m / z369-371-daughter ion m / z111-113;

[0023] The parent ion of the specific impurity is m / z355-357 and the product ion is m / z197-199;

[0024] Collision energy parameters:

[0025] Amisulpride is 20-30 eV;

[0026] Specific impurities are 25-35eV.

[0027] Preferably, in said S3, in the neutralizer solution, the histidine concentration is 8-12 g / L, the sodium thiosulfate concentration is 4-6 g / L, and the solvent is a phosphate buffer solution with a pH of 6.8-7.2.

[0028] Preferably, in said S5, the chromatography comprises:

[0029] The mobile phase is a mixture of methanol and 0.01-0.03 M ammonium acetate solution in a volume ratio of 35:65-45:55;

[0030] The detection wavelength is 225-235nm.

[0031] Preferably, in S4, flushing includes the following specific steps:

[0032] S401. After the test solution is filtered, add 25-35 mL of 0.1% peptone aqueous solution to the membrane filter, turn on the vacuum pump, maintain a negative pressure of -15 to -25 kPa, and allow the flushing solution to flow through the membrane within 10-15 seconds;

[0033] S402, add the flushing solution three times, each adding 100±5 mL, let it stand for 30-60 seconds after each addition, and then start vacuum negative pressure -20 to -30 kPa for filtration;

[0034] S403, add 50-70 mL of sterile pure water, filter under negative pressure, and then turn off the vacuum to keep the filter membrane moist;

[0035] The three flushing solutions are composed of:

[0036] First, 0.1% peptone aqueous solution;

[0037] the second, 0.1% peptone and 0.05% polysorbate 80 in water;

[0038] The third time, 0.1% peptone aqueous solution.

[0039] Preferably, the S5 further includes near-infrared rapid screening:

[0040] Collect oral solution at 11000-13000cm -1 and 3500-4500cm -1 Spectrum within the range;

[0041] The predicted value of amisulpride content was output through the partial least squares regression model.

[0042] Preferably, the test solution prepared in S3 is simultaneously used for S4 microbial limit test and S5 preservative test, wherein the operation interval between S4 and S5 is 0.5-2 hours, and the test solution is stored at 2-8°C during the interval.

[0043] The present invention provides a method for testing amisulpride oral solution. It has the following beneficial effects:

[0044] 1. The present invention uses the same test solution for both microbiological and chemical testing simultaneously, enabling two key analyses to be completed in a single preparation. Compared with the traditional method that requires processing two samples separately, this method completely solves the problem of system error accumulation caused by operational redundancy.

[0045] 2. The present invention unexpectedly forms a highly selective detection system through the unique combination of gradient program time control and collision energy range limitation. The existing method of independently optimizing chromatographic or mass spectrometric parameters is difficult to deal with the interference of complex degradation products. The present invention accurately identifies co-eluting impurities. This collaborative design provides a new path for monitoring trace risk substances.

[0046] 3. The present invention adopts a step-by-step targeted flushing process, combined with a proprietary neutralizer formula, to form a solution for completely removing antibacterial agents. Conventional methods result in fluctuations in microbial recovery rates due to insufficient neutralization. The present invention solves this problem through the synergistic effect of surfactant stage desorption and buffer components, demonstrating unique advantages in preparations containing sodium benzoate.

[0047] 4. The present invention integrates the acquisition of specific spectral bands into the detection process and establishes a real-time prediction model that does not require pre-processing. Compared with the limitations of traditional chromatographic detection, which is time-consuming and highly equipment-dependent, it achieves instant quality feedback during the production process and fills the gap in online monitoring of oral solutions. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 The present invention is a schematic flow chart of a method for testing an amisulpride oral solution. DETAILED DESCRIPTION

[0049] The following will clearly and completely describe the technical solution of the present invention in conjunction with the accompanying drawings. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0050] Please see the attached Figure 1 The embodiment of the present invention provides a method for testing an amisulpride oral solution, comprising the following steps:

[0051] S1. Sample pretreatment: The oral solution was activated, loaded, washed, and eluted through a solid phase extraction column in sequence, and the eluate was collected as the purification solution;

[0052] S2. Chromatography and mass spectrometry detection: The purified solution obtained in S1 is injected into an ultra-high performance liquid chromatography coupled with a triple quadrupole mass spectrometry system to detect the content of amisulpride and specific impurities under the set chromatographic and mass spectrometry conditions;

[0053] S3. Preparation of microbial test solution: Mix the oral solution with a neutralizer solution in a volume ratio, wherein the neutralizer comprises histidine and sodium thiosulfate;

[0054] S4. Microbial limit test: Take the test solution obtained in S3 and perform membrane filtration, washing and colony culture;

[0055] S5. Preservative detection: Take the test solution obtained in S3 and test the sodium benzoate content.

[0056] Specifically, the activation step removes impurities from the column, the elution separates the pigment and water-soluble excipients, and the elution selectively enriches the target. This process specifically addresses the interference of caramel pigment in the oral solution on the detection, ensuring that the purified liquid meets the requirements of ultra-trace detection.

[0057] In S1, activation, sample loading, washing, and elution include the following specific steps:

[0058] The activation operation uses 3-7 mL of methanol and 3-7 mL of pure water;

[0059] The elution operation is carried out using 3-7 mL of pure water and 3-7 mL of an aqueous solution containing 3-7% methanol;

[0060] The elution operation uses a mixed solvent of 3-7 mL of methanol and acetonitrile with a volume ratio of 1:0.8-1.2:1.4.

[0061] In S1, the eluate is concentrated and then diluted to 1.5-2.5 mL with 8-12 mM ammonium acetate solution containing 0.05-0.2% formic acid.

[0062] Specifically, the acidic reconstitution solution enhances the stability of mass spectrometry response, the buffer system maintains the ionization state of the target, the fixed volume is precisely adapted to the micro-volume injection system to avoid analyte loss during the concentration process, and the methanol-acetonitrile mixed elution system breaks through the limitations of traditional solvents and improves the desorption efficiency of the target.

[0063] In S2, the chromatographic conditions include:

[0064] Chromatographic column: C18 column with an inner diameter of 1.8-2.3 mm and a length of 80-120 mm, and a filler particle size of 1.5-2.0 μm;

[0065] Mobile phase A: 8-12 mM ammonium acetate solution containing 0.05-0.2% formic acid;

[0066] Mobile phase B contained 0.05-0.2% formic acid in acetonitrile.

[0067] Specifically, narrow-bore columns combined with sub-two-micron packings optimize mass transfer efficiency and shorten separation times for complex matrices. Acidic modification of the mobile phase suppresses peak tailing and improves chromatographic behavior reproducibility.

[0068] In S2, the mass spectrometry conditions set include:

[0069] Monitoring ion pair parameters:

[0070] Amisulpride parent ion m / z369-371-daughter ion m / z111-113;

[0071] The parent ion of the specific impurity is m / z355-357 and the product ion is m / z197-199;

[0072] Collision energy parameters:

[0073] Amisulpride is 20-30 eV;

[0074] Specific impurities are 25-35eV.

[0075] Specifically, dedicated ion pairs avoid interference from co-elution of excipients, and the dynamic range of collision energy adapts to instrument response variations. These two parameters synergistically ensure baseline separation of degradation products and the main component.

[0076] In S3, the neutralizer solution has a histidine concentration of 8-12 g / L, a sodium thiosulfate concentration of 4-6 g / L, and a solvent of phosphate buffer at pH 6.8-7.2.

[0077] Specifically, the histidine-sodium thiosulfate complex neutralizes antibacterial activity through a dual mechanism of action. It buffers pH to maintain the integrity of microbial cell membranes and overcomes the biocompatibility shortcomings of traditional neutralizers.

[0078] In S5, chromatography includes:

[0079] The mobile phase is a mixture of methanol and 0.01-0.03 M ammonium acetate solution in a volume ratio of 35:65-45:55;

[0080] The detection wavelength is 225-235nm.

[0081] Specifically, the mobile phase ratio optimizes the retention behavior of the preservative and avoids interference from the UV absorption of the main drug. The wavelength range covers the fluctuation of the detector's optical response, improving data acquisition stability.

[0082] In S4, flushing includes the following specific steps:

[0083] S401. After the test solution is filtered, add 25-35 mL of 0.1% peptone aqueous solution to the membrane filter, turn on the vacuum pump, maintain a negative pressure of -15 to -25 kPa, and allow the flushing solution to flow through the membrane within 10-15 seconds;

[0084] S402, add the flushing solution three times, each adding 100±5 mL, let it stand for 30-60 seconds after each addition, and then start vacuum negative pressure -20 to -30 kPa for filtration;

[0085] S403, add 50-70 mL of sterile pure water, filter under negative pressure, and then turn off the vacuum to keep the filter membrane moist;

[0086] The three flushing solutions are composed of:

[0087] First, 0.1% peptone aqueous solution;

[0088] the second, 0.1% peptone and 0.05% polysorbate 80 in water;

[0089] The third time, 0.1% peptone aqueous solution.

[0090] Specifically, in the step-by-step flushing strategy, the surfactant stage specifically removes the adsorption of the antibacterial agent, and the final step of pure water flushing maintains the osmotic pressure balance. The whole process solves the problem of microbial recovery rate attenuation caused by filter membrane residue.

[0091] S5 also includes near-infrared rapid screening:

[0092] Collect oral solution at 11000-13000cm -1 and 3500-4500cm -1 Spectrum within the range;

[0093] The predicted value of amisulpride content was output through the partial least squares regression model.

[0094] Specifically, characteristic band collection avoids interference from strong water absorption, and the chemometric model analyzes the spectral matrix effect. This solution enables non-destructive and rapid screening at the production site, breaking through the time bottleneck of traditional testing.

[0095] The test solution prepared in S3 is used for both S4 microbial limit test and S5 preservative test. The interval between S4 and S5 operations is 0.5-2 hours, and the test solution is stored at 2-8°C during the interval.

[0096] Specifically, a single test solution is reused across testing projects, and time control suppresses variation in sample properties. This design eliminates systematic bias introduced by repeated preparation and ensures the reliability of the correlation between microbial and chemical testing data.

[0097] The following is an introduction with reference to specific embodiments:

[0098] Example 1:

[0099] 2.0 mL of amisulpride oral solution was loaded onto a C18 solid-phase extraction column. Activation was performed using 5 mL of methanol and 5 mL of pure water at a flow rate of 1.5 mL / min. 5 mL of pure water and 5 mL of an aqueous solution containing 5% methanol were used for elution. 5 mL of a 1:1 volume ratio of methanol and acetonitrile was used for elution at a flow rate of 1.0 mL / min. The eluate was collected and concentrated to 0.3 mL under nitrogen blowdown at 40°C. The volume was then made up to 2.0 mL with 10 mM ammonium acetate solution containing 0.1% formic acid, and filtered through a 0.22 μm filter membrane.

[0100] Chromatography-mass spectrometry was performed on a 2.1×100 mm, 1.8 μm column. Mobile phase A consisted of 10 mM ammonium acetate in water containing 0.1% formic acid, and mobile phase B consisted of acetonitrile containing 0.1% formic acid. The gradient was 95% mobile phase A at 0 min, 80% at 3.0 min, 50% at 6.0 min, and 5% at 8.0 min. Mass spectrometry monitored the amisulpride parent ion m / z 370 corresponding to the daughter ion m / z 112 at a collision energy of 25 eV. The specific impurity parent ion m / z 356 corresponded to the daughter ion m / z 198 at a collision energy of 30 eV.

[0101] Preparation of microbial test solution: Dissolve 10 g / L histidine and 5 g / L sodium thiosulfate in pH 7.0 phosphate buffer to prepare a neutralizer. Take 10 mL of oral solution and mix with 100 mL of neutralizer at a ratio of 1:10.

[0102] Microbial limit test Take 1 mL of the test solution and filter it through a membrane. Rinse it by adding 30 mL of 0.1% peptone aqueous solution and filter under negative pressure for 12 seconds.

[0103] Add the rinse solution in three batches:

[0104] First, 100 mL of 0.1% peptone aqueous solution was allowed to stand for 45 seconds;

[0105] The second time, 100 mL of 0.1% peptone and 0.05% polysorbate 80 solution was allowed to stand for 45 s, and the third time, 100 mL of 0.1% peptone aqueous solution was allowed to stand for 45 s;

[0106] Finally, add 60 mL of sterile pure water and filter, split the filter membrane, and stick it on TSA plates and culture at 35℃ for 4 days and SDA plates at 25℃ for 6 days;

[0107] For preservative detection, 1.5 mL of the test solution was filtered and the mobile phase consisted of a mixture of methanol and 0.02 M ammonium acetate solution in a volume ratio of 40:60. The detection wavelength was 230 nm, the flow rate was 1.0 mL / min, and the column temperature was 30°C.

[0108] Table 1 shows the importance of simplifying the solid phase extraction and mass spectrometry detection process. In the prior art, solid phase extraction activation requires 3 steps and elution ≥ 3 times, which results in cumbersome operation and high reagent consumption. Mass spectrometry detection requires optimization of multiple groups of ion pairs, and method development is time-consuming. Unoptimized processes are prone to target loss and low detection efficiency. In contrast, Example 1 reduces the number of operating steps and the amount of reagents used by streamlining the activation step to 2 steps, combining the elution into 2 times, and standardizing the mass spectrometry monitoring parameters. This ensures efficient separation of amisulpride from impurities, ensures detection recovery, and shortens analysis time.

[0109] Example 2:

[0110] 1.5 mL of the oral solution was loaded. Activation was performed with 4 mL of methanol and 4 mL of pure water. Eluents were sequentially washed with 4 mL of pure water and 4 mL of a 7% methanol-water solution. Elution was performed with 4 mL of a 1:1.2 volume mixture of methanol and acetonitrile. The volume was then brought to 1.5 mL using 12 mM ammonium acetate solution containing 0.2% formic acid.

[0111] The specifications of the chromatographic column for chromatography-mass spectrometry detection were 2.1×100 mm 1.7 μm, and the gradient program was 98% of mobile phase A at 0 min, 85% at 3.5 min, 55% at 6.5 min, and 8% at 8.5 min.

[0112] The neutralizer for the preparation of the microbial test solution contains 12 g / L histidine and 6 g / L sodium thiosulfate, pH 7.2 phosphate buffer, with a mixing ratio of 1:12;

[0113] The second step of the main rinse for microbial limit testing uses a solution containing 0.1% peptone and 0.1% polysorbate 80. TSA medium is supplemented with 0.5% Tween 80;

[0114] The suitability of the preservative detection system is theoretical plate number ≥ 6000, and tailing factor 1.0.

[0115] Table 2 shows the importance of simplifying the chromatographic gradient and microbial reagents. In the prior art, the chromatographic gradient is segmented into ≥5 segments, and the microbial neutralizer contains 3 components, resulting in high mobile phase consumption, complex reagent preparation, and a large variety of culture medium additives that easily lead to component conflicts. The risk of contamination in microbial testing is high. In contrast, Example 2 reduces the gradient segmentation to 4 segments, simplifies the neutralizer components to 2, and unifies the culture medium additive to 0.5% Tween 80. This reduces mobile phase consumption, improves the repeatability of microbial testing, allows for rapid and complete testing, and reduces reagent costs.

[0116] Example 3:

[0117] The elution step was combined with 7 mL of pure water and 7 mL of an aqueous solution containing 3% methanol, and the elution was performed with 7 mL of a mixed solvent of methanol and acetonitrile in a volume ratio of 1:0.8, and the volume was adjusted to 2.5 mL with 8 mM ammonium acetate solution containing 0.05% formic acid;

[0118] The gradient program for chromatography-mass spectrometry detection was from 95% to 75% mobile phase A in the range of 0-2.5 min and from 75% to 50% in the range of 2.5-5.0 min, with a total run time of 8 min.

[0119] Preparation and testing of test solution: The same test solution should be tested for microorganisms first, and then tested for preservatives after an interval of 0.5 hours. The solution should be stored at 2°C during the interval.

[0120] Near infrared rapid screening directly takes undiluted oral solution and collects 12000-4000cm-1 Range spectra and content prediction using partial least squares regression model.

[0121] It can be seen from Table 3 that the importance of verifying the reuse of test solutions and rapid near-infrared detection is demonstrated. In the prior art, microbial and preservative detection require separate sample preparation, and content detection relies on complex instruments such as HPLC, resulting in large sample dosage and time-consuming detection, making it difficult to meet batch analysis requirements. The multi-item detection process in traditional methods is cumbersome and inefficient. In contrast, Example 3 uses the same test solution at intervals, combines near-infrared spectroscopy with the PLSR model, reduces sample dosage, shortens detection time, and improves scale detection efficiency.

[0122] Example 4:

[0123] A new elution step was added using 5 mL of 8% acetonitrile in water. Nitrogen purging was performed using high-purity nitrogen at 8 psi pressure.

[0124] The flow rate for the 6.5-8.5 min segment of the chromatographic mass spectrometry detection was set to 0.30 mL / min. The system suitability resolution was ≥ 2.2;

[0125] The microbial test solution was prepared by mixing in a biosafety cabinet and stored in sterile amber glass bottles.

[0126] Microbiological examination TSA medium was supplemented with 0.3% Tween 80, and SDA medium was supplemented with 0.03% chloramphenicol;

[0127] Near infrared screening spectral resolution 10cm -1 , using second-order derivative combined with multivariate scatter correction preprocessing.

[0128] From Table 4, we can know the importance of verifying the dynamic adjustment of chromatographic flow rate and the standardization of operating environment. In the prior art, constant flow throughout the chromatographic process can easily lead to peak overlap, non-standard microbial operating environment can easily cause contamination, and complex near-infrared spectral pretreatment affects the accuracy of the model. Unoptimized methods result in insufficient separation and low detection success rate. In contrast, Example 4 reduces the flow rate to 0.3mL / min in the 6.5-8.5 minute segment, and operates and stores in a biological safety cabinet in a sterile manner. Combined with the second-order derivative + MSC pretreatment spectrum, a separation degree ≥ 2.2 is achieved, the microbial contamination rate is reduced from 5% to 0.5%, and the near-infrared model prediction correlation R 2 Improved to 0.98 to ensure the stability and accuracy of the method.

[0129] Table 1: Standardization and simplification of solid phase extraction and mass spectrometry detection

[0130]

[0131] Table 2: Chromatographic gradients and reagents for microbial detection

[0132]

[0133]

[0134] Table 3: Test solution reuse and near-infrared rapid detection

[0135]

[0136]

[0137] Table 4: Detection parameters and operating environment

[0138]

[0139] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A method for testing amisulpride oral solution, characterized in that: The following steps are involved: S1. Sample pretreatment: The oral solution was activated, loaded, washed, and eluted through a solid phase extraction column in sequence, and the eluate was collected as the purification solution; S2. Chromatography and mass spectrometry detection: The purified solution obtained in S1 is injected into an ultra-high performance liquid chromatography coupled with a triple quadrupole mass spectrometry system to detect the content of amisulpride and specific impurities under the set chromatographic and mass spectrometry conditions; S3. Preparation of microbial test solution: mixing the oral solution and the neutralizer solution in a volume ratio to obtain a test solution, wherein the neutralizer comprises histidine and sodium thiosulfate; S4. Microbial limit test: Take the test solution obtained in S3 and perform membrane filtration, washing and colony culture; S5. Preservative detection: Take the test solution obtained in S3 and test the sodium benzoate content.

2. A method for testing amisulpride oral solution according to claim 1, characterized in that: In S1, activation, sample loading, washing and elution include the following specific steps: The activation operation uses 3-7 mL of methanol and 3-7 mL of pure water; The elution operation uses 3-7 mL of pure water and 3-7 mL of an aqueous solution containing 3-7% methanol in sequence; The elution operation uses a mixed solvent of 3-7 mL of methanol and acetonitrile, with a volume ratio of methanol to acetonitrile of 1:0.8-1.2:1.

4.

3. A method for testing amisulpride oral solution according to claim 1, characterized in that: In the above S1, the eluate is concentrated and then diluted to 1.5-2.5 mL with 8-12 mM ammonium acetate solution containing 0.05-0.2% formic acid, and then the content of amisulpride and specific impurities is detected in S2.

4. A method for testing amisulpride oral solution according to claim 1, characterized in that: In said S2, the chromatographic conditions include: Chromatographic column: C18 column with an inner diameter of 1.8-2.3 mm and a length of 80-120 mm, and a filler particle size of 1.5-2.0 μm; Mobile phase A: 8-12 mM ammonium acetate solution containing 0.05-0.2% formic acid; Mobile phase B contained 0.05-0.2% formic acid in acetonitrile.

5. A method for testing amisulpride oral solution according to claim 1, characterized in that: In said S2, the mass spectrometry conditions set include: Monitoring ion pair parameters: Amisulpride parent ion m / z369-371-daughter ion m / z111-113; The parent ion of the specific impurity is m / z355-357 and the product ion is m / z197-199; Collision energy parameters: Amisulpride is 20-30 eV; Specific impurities are 25-35eV.

6. A method for testing amisulpride oral solution according to claim 1, characterized in that: In the S3, the neutralizer solution has a histidine concentration of 8-12 g / L, a sodium thiosulfate concentration of 4-6 g / L, and a solvent of a phosphate buffer solution with a pH of 6.8-7.

2.

7. A method for testing amisulpride oral solution according to claim 1, characterized in that: In said S5, the chromatography method adopts the following steps: The mobile phase is a mixture of methanol and 0.01-0.03 M ammonium acetate solution in a volume ratio of 35:65-45:55; The detection wavelength is 225-235nm.

8. A method for testing amisulpride oral solution according to claim 1, characterized in that: In said S4, flushing comprises the following specific steps: S401. After the test solution is filtered, add 25-35 mL of 0.1% peptone aqueous solution to the membrane filter, turn on the vacuum pump, maintain a negative pressure of -15 to -25 kPa, and allow the flushing solution to flow through the membrane within 10-15 seconds; S402, add the flushing solution three times, each adding 100±5 mL, let it stand for 30-60 seconds after each addition, and then start vacuum negative pressure -20 to -30 kPa for filtration; S403, add 50-70 mL of sterile pure water, filter under negative pressure, and then turn off the vacuum to keep the filter membrane moist; The three flushing solutions are composed of: First, 0.1% peptone aqueous solution; the second, 0.1% peptone and 0.05% polysorbate 80 in water; The third time, 0.1% peptone aqueous solution.

9. A method for testing amisulpride oral solution according to claim 1, characterized in that: Said S5 also includes a near infrared rapid screening step: Collect oral solution at 11000-13000cm -1 and 3500-4500cm -1 Spectrum within the range; The predicted value of amisulpride content was output through the partial least squares regression model.

10. The method for testing amisulpride oral solution according to claim 1, wherein: The test solution prepared in S3 is simultaneously used for S4 microbial limit test and S5 preservative test, wherein the interval between the operations of S4 and S5 is 0.5-2 hours, and the test solution is stored at 2-8°C during the interval.

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