Detection method of compound sodium picosulfate preparation

Abstract

The invention relates to a detection method of a compound sodium picosulfate preparation, which is used for detecting sodium picosulfate and sodium benzoate in the compound sodium picosulfate preparation, the detection method adopts a high performance liquid chromatography, and the conditions of the high performance liquid chromatography are as follows: (1) a chromatographic column is a reverse C18 chromatographic column or an efficacy equivalent column; (2) the mobile phase A is a mixed solution of an ammonium acetate buffer solution and methanol, the pH value of the ammonium acetate buffer solution is 3.90-4.10, the volume percentage of the ammonium acetate buffer solution is 90%-98%, and the volume percentage of the methanol is 2%-10%; (3) a mobile phase B is acetonitrile; (4) isocratic elution is adopted, and the volume ratio of the mobile phase A to the mobile phase B is (87%-89%): (11%-13%). According to the detection method, sodium picosulfate and sodium benzoate in the compound sodium picosulfate preparation can be detected at the same time, and the detection accuracy is high.

Description

Technical Field

[0001] The present application relates to the technical field of preparation detection, and in particular to a detection method for a compound sodium picosulfate preparation. Background Art

[0002] Sodium picosulfate is a stimulant laxative that can be metabolized into bis(p-hydroxyphenyl)-pyridyl-2-methane (BHPM) by colonic bacteria. BHPM can effectively promote intestinal peristalsis. Compound sodium picosulfate granules contain magnesium oxide and citric acid. When magnesium oxide and citric acid meet water, they form magnesium citrate, which can effectively inhibit the loss of water with fecal excretion and act as an isotonic agent with zero water in the gastrointestinal tract. However, due to the difficulty of compound sodium picosulfate granules to dissolve quickly in water and generate a lot of heat, as well as the instability of citric acid and magnesium oxide in the air, people have further developed compound sodium picosulfate oral solution.

[0003] At present, the detection of sodium picosulfate in the pharmacopoeias of various countries is titration method. In addition, the content of sodium picosulfate in sodium picosulfate tablets is determined by high performance liquid chromatography, but this method is not suitable for the separation and detection of sodium picosulfate in compound sodium picosulfate preparations, and is prone to problems such as blocking the chromatogram. In addition, the existing technology does not involve the simultaneous detection of multiple components in compound sodium picosulfate preparations. Summary of the invention

[0004] Based on this, the present application provides a method for detecting a compound sodium picosulfate preparation that can simultaneously detect sodium picosulfate and sodium benzoate and has high detection accuracy.

[0005] The specific technical solutions are as follows:

[0006] A method for detecting a compound sodium picosulfate preparation is used to detect sodium picosulfate and sodium benzoate in the compound sodium picosulfate preparation. The detection method adopts high performance liquid chromatography, and the conditions of the high performance liquid chromatography include:

[0007] (1) The chromatographic column is a reversed C18 column or a column with equivalent performance;

[0008] (2) mobile phase A is a mixture of ammonium acetate buffer and methanol, wherein the pH of the ammonium acetate buffer is 3.90-4.10, the volume percentage of the ammonium acetate buffer is 90%-98%, and the volume percentage of methanol is 2%-10%;

[0009] (3) Mobile phase B is acetonitrile;

[0010] (4) Using isocratic elution, the volume ratio of mobile phase A to mobile phase B is (87% to 89%):(11% to 13%).

[0011] In one embodiment, the concentration of the ammonium acetate buffer is 25 mmol / L to 100 mmol / L.

[0012] In one embodiment, in the mobile phase A, the volume percentage of ammonium acetate buffer is 95%, and the volume percentage of methanol is 5%.

[0013] In one embodiment, the volume ratio of mobile phase A to mobile phase B is 88%:12%.

[0014] In one embodiment, the pH of the ammonium acetate buffer is 4.00; and / or,

[0015] The pH value of the ammonium acetate buffer was adjusted with glacial acetic acid.

[0016] In one embodiment, the column temperature of the HPLC is 43°C to 47°C.

[0017] In one embodiment, the conditions of the high performance liquid chromatography meet at least one of the following conditions:

[0018] (1) The detection wavelength is 260nm~265nm;

[0019] (2) Flow rate: 0.9 mL / min to 1.1 mL / min;

[0020] (3) Operation time ≥ 13 min;

[0021] (4) The injection volume is 5 μL to 15 μL.

[0022] In one embodiment, the reverse C18 chromatographic column is an Ultimate XB-C18 chromatographic column with a length of 150 mm, an inner diameter of 4.6 mm, and a filler particle size of 3 μm.

[0023] In one embodiment, the high performance liquid chromatography method using an external standard method to determine the content of sodium picosulfate and sodium benzoate in the compound sodium picosulfate preparation comprises the following steps:

[0024] preparing reference substance solutions of different concentrations, wherein the reference substance solutions contain sodium picosulfate and / or sodium benzoate;

[0025] The reference solution is tested by the high performance liquid chromatography method to prepare a standard curve;

[0026] According to the standard curve, calculating the content of sodium picosulfate and / or sodium benzoate in the compound sodium picosulfate preparation;

[0027] Wherein, in the reference solution, the concentration of sodium picosulfate is 20 μg / mL to 100 μg / mL; and / or, the concentration of sodium benzoate reference solution is 200 μg / mL to 900 μg / mL.

[0028] In one embodiment, the compound sodium picosulfate preparation includes a compound sodium picosulfate oral solution.

[0029] The detection method of the compound sodium picosulfate preparation can effectively separate sodium picosulfate and sodium benzoate in the preparation at the same time by adopting a specific type and ratio of mobile phase and isocratic elution, thereby eliminating the need to develop two detection conditions, reducing the detection cost, and solving the problem of chromatographic column blockage. In addition, the detection method has good specificity, good separation, high sensitivity, and good reproducibility, and can eliminate the interference caused by blank solvents and excipient solutions. It is highly practical in actual quality control work and is suitable for the quality control of the preparation process of the compound sodium picosulfate preparation and the final product. It is also of great significance for evaluating the quality of synthesis and preparation processes and stability sample testing. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 It is the blank solution chromatogram of the detection method of the compound sodium picosulfate preparation in the embodiment.

[0031] Figure 2 It is a chromatogram of the blank excipient solution of the detection method of the compound sodium picosulfate preparation in the embodiment.

[0032] Figure 3 It is a chromatogram of the reference substance solution of the detection method of the compound sodium picosulfate preparation in the example.

[0033] Figure 4 It is a chromatogram of the test solution of the detection method of the compound sodium picosulfate preparation in the embodiment.

[0034] Figure 5 The linearity test results of sodium picosulfate in the linearity investigation of the detection method of the compound sodium picosulfate preparation in the examples.

[0035] Figure 6 The linearity test results of sodium benzoate in the linearity investigation of the detection method of the compound sodium picosulfate preparation in the examples. DETAILED DESCRIPTION

[0036] The detection method of the compound sodium picosulfate preparation of the present application is further described in detail below in conjunction with specific examples. The present application can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present application more thorough and comprehensive.

[0037] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application belongs. The terms used herein in the specification of this application are only for the purpose of describing specific embodiments and are not intended to limit this application.

[0038] The optional scope of the terms "and / or", "or / and", and "and / or" used in this article includes any one of two or more related listed items, and also includes any and all combinations of the related listed items, said any and all combinations include any combination of any two related listed items, any more related listed items, or all related listed items.

[0039] As used herein, "one or more" refers to any one, any two, or any two or more of the listed items.

[0040] In the present application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0041] In this application, when it comes to numerical ranges, unless otherwise specified, the above numerical ranges are deemed to be continuous and include the minimum and maximum values ​​of the range, as well as each value between such minimum and maximum values. Further, when a range refers to an integer, each integer between the minimum and maximum values ​​of the range is included. In addition, when multiple ranges are provided to describe features or characteristics, the ranges can be merged. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges included therein.

[0042] The percentage contents involved in this application, unless otherwise specified, refer to mass percentage for solid-liquid mixing and solid-solid mixing, and refer to volume percentage for liquid-liquid mixing.

[0043] The percentage concentrations mentioned in this application, unless otherwise specified, refer to the final concentration, which refers to the percentage of the added component in the system after the addition of the component.

[0044] The temperature parameters in this application, unless otherwise specified, allow for both constant temperature treatment and treatment within a certain temperature range. The constant temperature treatment allows the temperature to fluctuate within the accuracy range of instrument control.

[0045] The room temperature in the present application generally refers to 4°C to 30°C, preferably 20±5°C.

[0046] Studies have shown that sodium picosulfate itself is easily soluble in water, has high polarity, and has a weak retention capacity in reversed-phase chromatography. If an acidic mobile phase is used, the more acidic it is, the less it will be retained. If an alkaline mobile phase is used, the OH in the mobile phase will -It is possible that Mg 2+ The reaction generates magnesium hydroxide precipitate, which blocks the chromatographic column, making it difficult to detect sodium picosulfate by reverse phase chromatography. At the same time, the applicant found that the compound sodium picosulfate preparation is easy to precipitate in the phosphate mobile phase, such as when the acetonitrile ratio exceeds 40%, the precipitate is precipitated and blocks the chromatographic column. It is speculated that the possible reason is the generation of magnesium phosphate precipitate; and the ammonium formate pka of the ammonium salt buffer is 3.75, and the buffer range is 2.75-4.75, which causes the sodium picosulfate to elute early and is difficult to separate from other substances in the compound sodium picosulfate preparation.

[0047] On this basis, the present application further develops a mobile phase elution program that matches it, which realizes the accurate detection of sodium picosulfate in the compound sodium picosulfate preparation, and can also simultaneously and accurately detect sodium benzoate in the compound sodium picosulfate preparation, solving the problems of being unable to simultaneously detect sodium picosulfate and sodium benzoate in the compound sodium picosulfate preparation, and chromatographic column blockage, which is conducive to comprehensive and efficient quality monitoring of the compound sodium picosulfate preparation.

[0048] In some examples of the present application, a method for detecting a compound sodium picosulfate preparation is provided, which is used to detect sodium picosulfate and sodium benzoate in the compound sodium picosulfate preparation. The detection method adopts high performance liquid chromatography, and the conditions of the high performance liquid chromatography include:

[0049] (1) The chromatographic column is a reversed C18 column or a column with equivalent performance;

[0050] (2) mobile phase A is a mixture of ammonium acetate buffer and methanol, wherein the pH of the ammonium acetate buffer is 3.90-4.10, the volume percentage of the ammonium acetate buffer is 90%-98%, and the volume percentage of methanol is 2%-10%;

[0051] (3) Mobile phase B is acetonitrile;

[0052] (4) Using isocratic elution, the volume ratio of mobile phase A to mobile phase B is (87% to 89%):(11% to 13%).

[0053] Specifically, the pH of the ammonium acetate buffer includes, but is not limited to, 3.90, 3.95, 4.00, 4.05, 4.10, or a range between any two of the foregoing. In mobile phase A, the volume percentage of the ammonium acetate buffer includes, but is not limited to, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or a range between any two of the foregoing; accordingly, the volume percentage of methanol includes, but is not limited to, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or a range between any two of the foregoing.

[0054] Specifically, the volume ratio of mobile phase A to mobile phase B includes, but is not limited to, 87%:13%, 88%:12%, 89%:11%, or a range between any two of the foregoing.

[0055] Without limitation, the isocratic elution time can be set to ≥ 13 min.

[0056] Without limitation, the pH of the ammonium acetate buffer can be adjusted by the addition of an acid, such as glacial acetic acid.

[0057] In some of these examples, the pH of the ammonium acetate buffer is 4.00.

[0058] In some of the examples, the volume percentage of the ammonium acetate buffer is 95% and the volume percentage of the methanol is 5%.

[0059] In some of the examples, the volume ratio of mobile phase A to mobile phase B is 88%:12%.

[0060] In some examples, the concentration of the ammonium acetate buffer is 25 mmol / L to 100 mmol / L. Specifically, the concentration of the ammonium acetate buffer includes but is not limited to: 25 mmol / L, 50 mmol / L, 75 mmol / L, 100 mmol / L, or a range between any two of the foregoing.

[0061] In some of the examples, the preparation method of the ammonium acetate buffer comprises weighing 1.9 g to 7.7 g of ammonium acetate and adding 1000 mL of water to dissolve it. Specifically, the mass of the ammonium acetate can be 1.9 g, 3.8 g, 5.7 g, or 7.7 g.

[0062] It is understood that "C18" means that the chromatographic column uses octadecylsilane bonded silica gel as a filler. Without limitation, the chromatographic column is Ultimate XB-C18 or a chromatographic column with equivalent performance. Further, the specifications of the chromatographic column include: length 140mm ~ 160mm, inner diameter 4mm ~ 5mm, filler particle size 2μm ~ 5μm. In some examples, the reverse C18 chromatographic column is Ultimate XB-C18 chromatographic column, length 150mm, inner diameter 4.6mm, filler particle size 3μm.

[0063] In some examples, the conditions of the high performance liquid chromatography method further include: a detection wavelength of 260 nm to 265 nm. Specifically, the detection wavelength includes but is not limited to: 260 nm, 261 nm, 262 nm, 263 nm, 264 nm, 265 nm, or a range between any two of the foregoing.

[0064] In some examples, the conditions of the high performance liquid chromatography method further include: a column temperature of 43° C. to 47° C. Specifically, the column temperature includes but is not limited to: 43° C., 44° C., 45° C., 46° C., 47° C., or a range between any two of the foregoing.

[0065] In some examples, the conditions of the high performance liquid chromatography method further include: a flow rate of 0.9 mL / min to 1.1 mL / min. Specifically, the flow rate includes but is not limited to: 0.9 mL / min, 1 mL / min, 1.1 mL / min, or a range between any two of the foregoing.

[0066] In some examples, the conditions of the high performance liquid chromatography method also include: the injection volume is 5 μL to 15 μL. Specifically, the flow rate includes but is not limited to: the injection volume includes but is not limited to: 5 μL, 8 μL, 10 μL, 12 μL, 15 μL, 15 μL, or a range between any two of the foregoing.

[0067] In some of these examples, the HPLC method using an external standard method to determine the content of sodium picosulfate and / or sodium benzoate in a compound sodium picosulfate preparation includes the following steps:

[0068] preparing reference solutions of different concentrations, wherein the reference solutions contain sodium picosulfate and / or sodium benzoate;

[0069] The reference solution was tested by high performance liquid chromatography to prepare a standard curve;

[0070] According to the standard curve, calculate the content of sodium picosulfate and / or sodium benzoate in the compound sodium picosulfate preparation.

[0071] In some of the examples, the concentration of sodium picosulfate in the reference solution is 20 μg / mL to 100 μg / mL. It is understandable that the concentrations of sodium picosulfate in the reference solutions of different concentrations are different, such as preparing 5 concentrations of reference solutions, where the concentrations of sodium picosulfate can be selected from the following concentrations: 20 μg / mL, 23 μg / mL, 46 μg / mL, 50 μg / mL, 57 μg / mL, 69 μg / mL, 70 μg / mL, 92 μg / mL, 100 μg / mL.

[0072] Similarly, in some of the examples, the concentration of sodium benzoate in the reference solution is distributed between 200 μg / mL and 900 μg / mL. The concentrations of sodium benzoate in the reference solutions of different concentrations are different. For example, 5 reference solutions of different concentrations are prepared, in which the concentrations of sodium benzoate can be selected from the following concentrations: 200 μg / mL, 205 μg / mL, 300 μg / mL, 400 μg / mL, 410 μg / mL, 500 μg / mL, 512 μg / mL, 600 μg / mL, 615 μg / mL, 700 μg / mL, 800 μg / mL, and 820 μg / mL.

[0073] It should be noted that a reference solution may contain only sodium picosulfate or sodium benzoate, or both sodium picosulfate and sodium benzoate, and the specific preparation can be based on actual conditions.

[0074] In addition, without limitation, the compound sodium picosulfate preparation includes a compound sodium picosulfate oral solution, and the prescription of the compound sodium picosulfate oral solution includes sodium picosulfate and / or sodium benzoate. Without limitation, when the compound sodium picosulfate preparation includes a compound sodium picosulfate oral solution, the preparation method of the test solution may include filtering the compound sodium picosulfate oral solution and obtaining a filtrate. The filter membrane used for filtration may be, for example, a PES filter membrane (0.45 μm).

[0075] It is understandable that the compound sodium picosulfate oral solution is shaken before filtering to improve the accuracy of the result. In addition, in order to reduce the detection error, the compound sodium picosulfate oral solution can be filtered using a PES filter membrane (0.45 μm), 1 mL of the initial filtrate is discarded, and then the test solution is obtained.

[0076] For experimental parameters not specified in the following specific embodiments, reference is made to the instructions given in the present application document, and reference may also be made to experimental manuals in the art or other experimental methods known in the art, or to experimental conditions recommended by manufacturers.

[0077] The raw materials and reagents involved in the following specific examples can be obtained from commercial sources, or can be prepared by those skilled in the art according to known methods.

[0078] 1. Instruments and reagents:

[0079] 1. Main instruments

[0080] Instrument: Waters Arc-2998 series high performance liquid chromatograph.

[0081] Chromatographic column: Ultimate XB-C18 4.6*150mm, 3μm.

[0082] Electronic balance: GL224I-1SCN (Sartorius).

[0083] pH meter: FE28 (Mettler).

[0084] 2. Main reagents

[0085] Ammonium acetate (analytical grade), glacial acetic acid (analytical grade), methanol (chromatographic grade), acetonitrile (chromatographic grade).

[0086] 3. Reference substances

[0087] Sodium picosulfate reference substance: Source: Bemi Research and Development (Shenzhen) Co., Ltd., content 96.7%, batch number: PIC-222019.

[0088] Sodium benzoate reference substance: sourced from China National Institute for Food and Drug Control, content 99.8%, batch number 100433-202103.

[0089] 4. Samples

[0090] Test sample: Compound sodium picosulfate oral solution, source: Huayi Pharmaceutical Technology (Anhui) Co., Ltd., content specification: 175mL, sodium picosulfate 10mg, excipients include sodium benzoate, citric acid, magnesium oxide, disodium edetate, sucralose, sodium metabisulfite, glycine, alanine, acesulfame potassium and sodium hydroxide.

[0091] Blank excipient: that is, compound sodium picosulfate oral solution that does not contain sodium picosulfate and sodium benzoate, provided by Bemi Research and Development (Shenzhen) Co., Ltd.

[0092] 2. Solution Preparation

[0093] 1. Blank solution: purified water.

[0094] 2. Diluent: Take appropriate amount of acetonitrile and water and mix them in the ratio of 8:92 (V / V).

[0095] 3. Sodium benzoate reference substance stock solution: Take about 20.5 mg of sodium benzoate reference substance (calculated as C7H5NaO2), weigh accurately, place in a 20 mL volumetric flask, add appropriate amount of diluent, dissolve by ultrasonication and dilute to scale, shake well and prepare two in parallel.

[0096] 4. Sodium picosulfate reference substance stock solution: weigh about 23 mg of sodium picosulfate reference substance (calculated according to C 18 H 13 100mL volumetric flask, dissolve with ultrasonic diluent and dilute to scale, shake well, and prepare two portions in parallel.

[0097] 5. Reference solution: Accurately measure 5 mL of sodium picosulfate reference solution and 10 mL of sodium benzoate reference solution, place in a 20 mL volumetric flask, add water to dilute to the mark, shake well, and prepare two portions in parallel. (Concentration of sodium picosulfate: 0.0575 mg / mL; concentration of sodium benzoate: 0.5125 mg / mL).

[0098] 6. Test solution: shake the test sample evenly, filter it through a 0.45 μm PES filter membrane, discard 1 mL of the initial filtrate, and take the subsequent filtrate as the test solution.

[0099] 7. Blank excipient solution: Take the blank excipient and shake it evenly, filter it with a 0.45μm PES filter membrane, discard 1mL of the initial filtrate, and take the subsequent filtrate as the blank excipient solution.

[0100] 3. Determination methods and conditions

[0101] A method for detecting a compound sodium picosulfate preparation is used to detect sodium picosulfate and sodium benzoate. The detection method adopts high performance liquid chromatography. The conditions of the high performance liquid chromatography are shown in Table 1 below:

[0102] Table 1 HPLC detection conditions of compound sodium picosulfate preparation

[0103]

[0104] 4. Calculation formula

[0105] 1. External standard method:

[0106]

[0107]

[0108] in,

[0109] M st The sample weight of the reference substance (mg);

[0110] P st is the content of reference substance (%);

[0111] V st is the dilution volume of the reference substance, mL;

[0112] A st is the peak area of ​​sodium picosulfate and sodium benzoate in the reference solution;

[0113] A s is the peak area of ​​sodium picosulfate and sodium benzoate in the test solution;

[0114] n s is the dilution multiple of the test product;

[0115] is the average F value of reference solution 1 (n=5);

[0116] V is the finished product packaging volume, 175mL;

[0117] L is the labeled amount of sodium picosulfate: 10 mg; the labeled amount of sodium benzoate: 90 mg.

[0118] 2. Recovery rate:

[0119] Recovery rate = measured amount / added amount × 100%.

[0120] 3.Relative standard deviation (RSD):

[0121]

[0122] Where S is the standard deviation, It is the arithmetic mean of the measurement results.

[0123] V. Specific test content

[0124] Example 1

[0125] A method for detecting a compound sodium picosulfate preparation is used to detect sodium picosulfate and sodium benzoate in the compound sodium picosulfate preparation. The method adopts high performance liquid chromatography. The conditions of the high performance liquid chromatography include:

[0126] Chromatographic column: Ultimate XB-C18, 4.6*150mm, 3μm;

[0127] Mobile phase: Phase A: ammonium acetate buffer-methanol 95:5, Phase B: acetonitrile; Phase A: Phase B is 88:12 (V / V); wherein the preparation method of the ammonium acetate buffer comprises: weighing 7.7 g of ammonium acetate, adding 1000 mL of water to dissolve, and the concentration of the ammonium acetate buffer is 100 mmol / L; adjusting the pH to 4.00 with glacial acetic acid;

[0128] Elution mode: isocratic elution;

[0129] Flow rate: 1.0 mL / min;

[0130] Column temperature: 45°C;

[0131] Wavelength: 263nm;

[0132] Injection volume: 10 μL;

[0133] Running time: 15 minutes;

[0134] The reference solution and the test solution were respectively injected into the HPLC, and the contents of sodium picosulfate and sodium benzoate were calculated by the external standard method.

[0135] Example 2

[0136] Compared with Example 1, the column temperature was adjusted to 43° C., and other chromatographic conditions remained unchanged.

[0137] Example 3

[0138] Compared with Example 1, the column temperature was adjusted to 47° C., and other chromatographic conditions remained unchanged.

[0139] Example 4

[0140] Compared with Example 1, the flow rate was adjusted to 0.9 mL / min, and other chromatographic conditions remained unchanged.

[0141] Example 5

[0142] Compared with Example 1, the flow rate was adjusted to 1.1 mL / min, and other chromatographic conditions remained unchanged.

[0143] Example 6

[0144] Compared with Example 1, the pH of the ammonium acetate buffer was adjusted to 3.90 with glacial acetic acid, and other chromatographic conditions remained unchanged.

[0145] Example 7

[0146] Compared with Example 1, the pH of the ammonium acetate buffer was adjusted to 4.10 with glacial acetic acid, and other chromatographic conditions remained unchanged.

[0147] Example 8

[0148] Compared with Example 1, the concentration of ammonium acetate buffer was 50 mmol / L, and other chromatographic conditions remained unchanged.

[0149] Example 9

[0150] Compared with Example 1, the concentration of ammonium acetate buffer was 25 mmol / L, and other chromatographic conditions remained unchanged.

[0151] Example 10

[0152] Compared with Example 1, in mobile phase A, the ratio of ammonium acetate buffer to methanol was 98-2; the ratio of mobile phase A to mobile phase B was 87:13 (V / V), and other chromatographic conditions remained unchanged.

[0153] Embodiment 11

[0154] Compared with Example 1, in mobile phase A, the ratio of ammonium acetate buffer to methanol is 90-10; the ratio of mobile phase A to mobile phase B is 89:11 (V / V), and other chromatographic conditions remain unchanged.

[0155] Comparative Example 1

[0156] Compared with Example 1, the column temperature was 30°C, the injection volume was 10 μL, the mobile phase A was 100 mmol / L ammonium acetate buffer, and the pH was adjusted to 3.80 with glacial acetic acid; the mobile phase B was acetonitrile, and the ratio of mobile phase A to mobile phase B was 82.5:17.5 (V / V), and the other chromatographic conditions remained unchanged.

[0157] Comparative Example 2

[0158] Compared with Example 1, the column temperature was 30°C, the injection volume was 20 μL, the mobile phase A was 25 mmol / L ammonium acetate buffer, and the pH was adjusted to 4.20 with glacial acetic acid; the mobile phase B was acetonitrile, and the ratio of mobile phase A to mobile phase B was 85:15 (V / V), and the other chromatographic conditions remained unchanged.

[0159] Comparative Example 3

[0160] Compared with Example 1, mobile phase A was 100 mmol / L ammonium acetate buffer (without pH adjustment), mobile phase B was acetonitrile, and other chromatographic conditions remained unchanged.

[0161] The test results of the test solution of this embodiment and the comparative example are shown in Table 2, and the chromatograms of the blank solution, blank auxiliary material, reference solution and test solution of Example 1 are shown in Figures 1 to 4 .

[0162] Table 2 Data of test solution of each embodiment and comparative example

[0163]

[0164] As shown in Table 2, Examples 1 to 11 can achieve effective separation of sodium picosulfate and sodium benzoate in the test sample by using different HPLC conditions within the scope of the present application, and then achieve accurate detection of the two, and there is no chromatographic column clogging. However, Comparative Examples 1 to 3 use HPLC conditions different from those of the present application, and all have problems such as impurity peak interference, poor peak shape, and inability to detect simultaneously, making it difficult to achieve effective separation and accurate detection of the two.

[0165] VI. Method Validation Content

[0166] The determination method of Example 1 was used for methodological verification to verify its feasibility and stability.

[0167] 1. System adaptability

[0168] Two reference solutions were prepared in parallel, respectively counted as reference solution 1 and reference solution 2, and the chromatographic conditions of Example 1 were used to detect in the following order: 5 injections of reference solution 1, 2 injections of reference solution 2, and 1 injection of reference solution 1, and a PDA detector was selected to record the chromatogram. The acceptable standards and results are shown in Table 3 below.

[0169] Table 3 System suitability results

[0170]

[0171] Note: N / A means not applicable.

[0172] From the above results, it can be seen that the reference solution 1 was injected continuously for 5 times, and the RSDs of the peak areas of sodium picosulfate and sodium benzoate were 0.3% and 0.1%, respectively; the recoveries of sodium picosulfate and sodium benzoate of the reference solution 2 relative to the reference solution 1 were 100.0% and 100.7%, respectively; the RSDs (n=6) of the peak areas of sodium picosulfate and sodium benzoate of the Huizhen solution and the reference solution 1 were 0.3% and 0.2%, respectively, which were all in compliance with the regulations; therefore, the system applicability of this method meets the requirements.

[0173] 2. Accuracy

[0174] Reference substance stock solution: Take 23 mg of sodium picosulfate reference substance and 205 mg of sodium benzoate reference substance, accurately weigh, place in a 100 mL volumetric flask, add diluent, ultrasonically dissolve and dilute to the scale, shake well, and obtain.

[0175] 40% accuracy solution: accurately measure 2 mL of reference stock solution, place in a 20 mL volumetric flask, quantitatively dilute to the scale with blank excipient solution, shake well, filter with a 0.45 μm PES filter membrane, discard 1 mL of the initial filtrate, and obtain the subsequent filtrate; prepare 3 copies in parallel.

[0176] 100% accuracy solution: accurately measure 5 mL of reference stock solution, place in a 20 mL volumetric flask, quantitatively dilute to the mark with blank excipient solution, shake well, filter with a 0.45 μm PES filter membrane, discard 1 mL of the initial filtrate, and obtain the subsequent filtrate; prepare 3 copies in parallel.

[0177] 160% accuracy solution: Accurately measure 4 mL of the reference linear stock solution (see the "Linearity and Range" below for details), place it in a 10 mL volumetric flask, quantitatively dilute it to the scale with the blank excipient solution, shake well, filter it with a 0.45 μm PES filter membrane, discard 1 mL of the initial filtrate, and take the subsequent filtrate; prepare 3 copies in parallel.

[0178] Take 10 μL of each of the above solutions, inject into the liquid chromatograph, record the chromatogram, and calculate the recovery rate and RSD of each solution by the external standard method. The acceptable standards and results are shown in Table 4 below.

[0179] Table 4 Accuracy test results

[0180]

[0181] It can be seen from the data results that when sodium picosulfate was in the range of 0.0231 mg / mL to 0.0923 mg / mL (equivalent to 40.4% to 161.6% of the limit concentration), the recovery rate of sodium picosulfate in 9 test solutions was between 99.7% and 100.9%, the average recovery rate was 100.2%, and the RSD was 0.4%; when sodium benzoate was in the range of 0.2052 mg / mL to 0.8208 mg / mL (equivalent to 39.9% to 159.6% of the limit concentration), the recovery rate of sodium picosulfate in 9 test solutions was between 100.1% and 101.0%, the average recovery rate was 100.5%, and the RSD was 0.4%; all met the requirements, and the accuracy results of sodium picosulfate and sodium benzoate met the requirements.

[0182] 3. Filter membrane adsorption

[0183] Centrifugal solution: Shake the test sample, centrifuge (10000rpm, 5min), and take the supernatant as the centrifugal solution.

[0184] Membrane adsorption test solution: Use PES filter membrane for the test sample (two filter membranes, PES-1 and PES-2, are tested in parallel), discard 0mL, 1mL, and 2mL of the initial filtrate, and take the subsequent filtrate.

[0185] Take 10 μL of each of the above solutions, inject into the liquid chromatograph, record the chromatogram, and compare the peak areas of sodium picosulfate and sodium benzoate of the membrane adsorption test solution and the reference solution. The acceptable standards and results are shown in Tables 5 and 6 below:

[0186] Table 5 Sodium picosulfate membrane adsorption test results

[0187]

[0188] Note: NA means not applicable.

[0189] Table 6 Sodium benzoate membrane adsorption test results

[0190]

[0191] Note: NA means not applicable.

[0192] From the above, it can be seen that when 0mL, 1mL, and 2mL of the primary filtrate are discarded using a PES filter membrane (0.45μm), in the filtered test solution, the ratio of the peak area of ​​sodium picosulfate to the peak area of ​​the sodium picosulfate peak in the centrifuged solution is between 0.99 and 1.00; the ratio of the peak area of ​​sodium benzoate to the peak area of ​​sodium benzoate in the centrifuged solution is 1.00, both of which are between 0.98 and 1.02, which meets the requirements. Therefore, this product is filtered with a PES filter membrane (0.45μm) and 1mL of the primary filtrate is discarded.

[0193] 4. Repeatability

[0194] Prepare 6 test sample solutions, inject 10 μL of each solution into the liquid chromatograph, record the chromatogram, and calculate the content and RSD of the 6 test samples by the external standard method. The acceptable standards and results are shown in Table 7 below:

[0195] Table 7 Repeatability test results

[0196]

[0197] From the above, it can be seen that the average content of sodium picosulfate in the 6 test solutions is 101.1% and the RSD is 0.2%; the average content of sodium benzoate is 100.2% and the RSD is 0.2%, both of which meet the requirements and the repeatability meets the requirements.

[0198] 5. Reproducibility

[0199] Two people prepared the solutions according to the solution preparation method, and injected 10 μL of each solution into the liquid chromatograph to record the chromatogram. The acceptable standards and results are shown in Table 8:

[0200] Table 8 Reproducibility test results

[0201]

[0202]

[0203] From the above, we can know that the RSD of the content of sodium picosulfate in 6 test solutions is 0.2% and 0.09%, which is in compliance with the regulations, and the RSD of the content of sodium picosulfate in 12 test solutions of two experimenters is 0.6%, which is in compliance with the regulations; the RSD of the content of sodium benzoate in 6 test solutions is 0.2% and 0.05%, which is in compliance with the regulations, and the RSD of the content of sodium benzoate in 12 test solutions of two experimenters is 1.1%, which is in compliance with the regulations. Therefore, the reproducibility meets the requirements.

[0204] 6. Linearity and range

[0205] Linear stock solution: Take 23 mg of sodium picosulfate reference substance and 205 mg of sodium benzoate reference substance, accurately weigh, place in a 100 mL volumetric flask, add diluent, ultrasonically dissolve and dilute to the scale, shake well, and obtain.

[0206] 40% linear solution: Accurately measure 2 mL of linear stock solution, place in a 20 mL volumetric flask, quantitatively dilute to the scale with water, shake well, and obtain.

[0207] 80% linear solution: Accurately measure 2 mL of linear stock solution, place it in a 10 mL volumetric flask, quantitatively dilute it to the scale with water, shake well, and obtain.

[0208] 100% linear solution: Accurately measure 5 mL of linear stock solution, place it in a 20 mL volumetric flask, quantitatively dilute it to the scale with water, shake well, and obtain.

[0209] 120% linear solution: Accurately measure 3 mL of linear stock solution, place it in a 10 mL volumetric flask, quantitatively dilute it to the scale with water, shake well, and obtain.

[0210] 160% linear solution: Accurately measure 4 mL of linear stock solution, place in a 10 mL volumetric flask, quantitatively dilute to the scale with water, shake well, and obtain.

[0211] Accurately measure 10 μL of each of the above solutions, inject into the liquid chromatograph, and record the chromatogram. With concentration as the horizontal axis and peak area as the vertical axis, calculate the regression equation and correlation coefficient by the least squares method, the intercept is equivalent to the percentage of the limit concentration response, and the residual sum of squares.

[0212] The linear test results are shown in Tables 9 and 10. Figure 5 and Figure 6 As shown, the acceptance criteria are: the correlation coefficient of the linear regression equation r ≥ 0.995, and the ratio of the reported Y-axis intercept to the limit concentration content response value should not be greater than 3.0%:

[0213] Table 9 Sodium picosulfate linearity test results

[0214]

[0215] Table 10 Sodium benzoate linearity test results

[0216]

[0217] From the above data, it can be seen that the linear regression equation of sodium picosulfate is y=7160.6890x+2078.8500, r=1.0000, the ratio of the y-axis intercept to the limit concentration response value is 0.5%, and the residual sum of squares is 8567931, indicating that sodium picosulfate has a good linear relationship in the range of 23.08μg / mL to 92.33μg / mL (equivalent to a limit concentration of 40.4% to 161.6%); the linear regression equation of sodium benzoate is y=2326.5368x+4678.1500, r=1.0000, the ratio of the y-axis intercept to the limit concentration response value is 0.4%, and the residual sum of squares is 49972749, indicating that sodium benzoate has a good linear relationship in the range of 205.19μg / mL to 820.76μg / mL (equivalent to 39.9% to 159.6% of the limit concentration).

[0218] 7. Solution stability

[0219] Place the reference solution and the test solution at room temperature and away from light, take 10 μL of the above solution for injection analysis at different time points (e.g. 0 h, 7.75 h, 37 h, 74.5 h, 76.6 h), and record the chromatogram.

[0220] The specific inspection time points were adjusted according to the actual injection time, and the peak area ratio of each time point to 0 hour or the content ratio of each time point to 0 hour was calculated.

[0221] The acceptance criteria and results are shown in Table 11 below.

[0222] Table 11 contains solution stability test results

[0223]

[0224]

[0225] Note: NA means not applicable.

[0226] As can be seen from the above, the system suitability at each time point meets the requirements. When placed at room temperature and away from light, the peak area ratios of sodium picosulfate and sodium benzoate of the reference solution within 76.6 hours to the peak area at 0 hours are both between 1.00 and 1.01, which meets the requirements; the peak area ratios of sodium picosulfate and sodium benzoate of the test solution within 74.5 hours to the peak area at 0 hours are 0.99 to 1.00, respectively, which also meets the requirements. This indicates that the reference solution can be used at least within 76.6 hours at room temperature and away from light, and the test solution can be used at least within 74.5 hours at room temperature and away from light.

[0227] In summary, the system adaptability, specificity, accuracy, repeatability, reproducibility, linearity and range of the detection method of the present application all meet the requirements, and can effectively separate and accurately quantify sodium picosulfate and sodium benzoate in compound sodium picosulfate oral solution at the same time, and the method has good specificity, good separation and good accuracy. In addition, the stability of the test solution and the reference solution is also investigated, and the results show that both are stable within at least 74.5h. Therefore, it is explained that the detection method of the present application can be used for the daily detection of the sodium picosulfate and sodium benzoate content of the compound sodium picosulfate oral solution, the test sample preparation process detection, etc.

[0228] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0229] The above-described embodiments only express several implementation methods of the present application, which is convenient for understanding the technical solution of the present application in detail, but it cannot be understood as limiting the scope of protection of the patent application. It should be pointed out that for ordinary technicians in this field, without departing from the concept of the present application, several modifications and improvements can be made, which all belong to the scope of protection of the present application. It should be understood that the technical solutions obtained by those skilled in the art through logical analysis, reasoning or limited experiments on the basis of the technical solutions provided in the present application are all within the scope of protection of the claims attached to the present application. Therefore, the scope of protection of the patent of this application shall be based on the content of the attached claims, and the description can be used to interpret the content of the claims.

Claims

1. A method for detecting a compound sodium picosulfate preparation, characterized in that: Used to detect sodium picosulfate and sodium benzoate in a compound sodium picosulfate preparation, the detection method adopts high performance liquid chromatography, and the conditions of the high performance liquid chromatography include: (1) The chromatographic column is a reversed C18 column or a column with equivalent performance; (2) mobile phase A is a mixture of ammonium acetate buffer and methanol, wherein the pH of the ammonium acetate buffer is 3.90-4.10, the volume percentage of the ammonium acetate buffer is 90%-98%, and the volume percentage of methanol is 2%-10%; (3) mobile phase B is acetonitrile; (4) Using isocratic elution, the volume ratio of mobile phase A to mobile phase B is (87% to 89%):(11% to 13%).

2. The detection method of the compound sodium picosulfate preparation according to claim 1, characterized in that: The concentration of the ammonium acetate buffer is 25 mmol / L to 100 mmol / L.

3. The detection method of the compound sodium picosulfate preparation according to claim 2, characterized in that: In the mobile phase A, the volume percentage of ammonium acetate buffer is 95%, and the volume percentage of methanol is 5%.

4. The detection method of the compound sodium picosulfate preparation according to claim 3, characterized in that: The volume ratio of mobile phase A to mobile phase B was 88%:12%.

5. The detection method of the compound sodium picosulfate preparation according to claim 4, characterized in that: The pH of the ammonium acetate buffer is 4.00; and / or, The pH value of the ammonium acetate buffer was adjusted with glacial acetic acid.

6. The method for detecting the compound sodium picosulfate preparation according to any one of claims 1 to 5, characterized in that: The column temperature of the high performance liquid chromatography is 43°C to 47°C.

7. The detection method of the compound sodium picosulfate preparation according to any one of claims 1 to 5, characterized in that: The conditions of the high performance liquid chromatography method meet at least one of the following conditions: (1) The detection wavelength is 260nm~265nm; (2) Flow rate: 0.9 mL / min to 1.1 mL / min; (3) Operation time ≥ 13 min; (4) The injection volume is 5 μL to 15 μL.

8. The method for detecting the compound sodium picosulfate preparation according to any one of claims 1 to 5, characterized in that: The reverse C18 chromatographic column is an Ultimate XB-C18 chromatographic column, which is 150 mm long, 4.6 mm in inner diameter, and has a filler particle size of 3 μm.

9. The method for detecting the compound sodium picosulfate preparation according to any one of claims 1 to 5, characterized in that: The high performance liquid chromatography method adopts an external standard method to determine the content of sodium picosulfate and sodium benzoate in the compound sodium picosulfate preparation, comprising the following steps: preparing reference substance solutions of different concentrations, wherein the reference substance solutions contain sodium picosulfate and / or sodium benzoate; The reference solution is tested by the high performance liquid chromatography method to prepare a standard curve; According to the standard curve, calculating the content of sodium picosulfate and / or sodium benzoate in the compound sodium picosulfate preparation; Wherein, in the reference solution, the concentration of sodium picosulfate is 20 μg / mL to 100 μg / mL; and / or, the concentration of sodium benzoate reference solution is 200 μg / mL to 900 μg / mL.

10. The method for detecting the compound sodium picosulfate preparation according to any one of claims 1 to 5, characterized in that: The compound sodium picosulfate preparation comprises a compound sodium picosulfate oral solution.

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