Method for simultaneously determining contents of sorbitol, xylitol, sucralose and sodium benzoate

Abstract

The invention discloses a method for simultaneously determining the contents of sorbitol, xylitol, sucralose and sodium benzoate in a solution, which comprises the following steps of: taking a proper amount of solution, diluting with water, and taking centrifuged supernate as a test solution; and determining the contents of sorbitol, xylitol, sucralose and sodium benzoate in the test solution by using a high performance liquid chromatography-mass spectrometer, and comparing the contents with labeled amounts. According to the method for determining the contents of sorbitol, xylitol, sucralose and sodium benzoate, the quality of samples among batches of preparations can be evaluated, indexes are provided for research and development of oral solutions, a guarantee is provided for optimization of a preparation process, and the consistency of the quality and the curative effect of a medicine can be confirmed after a product is changed to some extent.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical analysis and detection, and particularly to a method for simultaneously determining the contents of sorbitol, xylitol, sucralose and sodium benzoate in a solution preparation. Background Art

[0002] The chemical name of sorbitol in the present invention is (2R,3R,4R,5S)-hexane-1,2,3,4,5,6-hexol, and its chemical formula is C 6 H 14 O 6 , and its chemical structural formula is as shown in (I), with a molecular weight of 182.18; the chemical name of xylitol is (2S,4R)-pentane-1,2,3,4,5-pentol, and its chemical formula is C 5 H 12 O 5 , and its chemical structural formula is as shown in (II), with a molecular weight of 152.15; the chemical name of sucralose is (2R,3R,4R,5R,6R)-2-[(2R,3S,4S,5S)-2,5-bis(chloromethyl)-3,4-dihydroxyoxolan-2-yl]oxy-5-chloro-6-(hydroxymethyl)oxane-3,4-diol, and its chemical formula is C 12 H 19 Cl 3 O 8 , and its chemical structural formula is as shown in (III), with a molecular weight of 397.63. The chemical name of sodium benzoate is sodium benzoate, and its chemical formula is C 7 H 5 NaO 2 , and its chemical structural formula is as shown in (IV), with a molecular weight of 144.10.

[0003]

[0004] Sorbitol, xylitol, and sucralose are commonly used sweeteners in oral solutions, and sodium benzoate is a commonly used bacteriostatic agent in oral solutions. Their maximum daily exposure amounts in the FDA excipient database are 24,000 mg, 21,600 mg, 192 mg, and 120 mg respectively. The dosage ratios of sorbitol, xylitol, sucralose, and sodium benzoate are the main research contents in the formulation of oral solutions. Currently, no relevant methods for the determination of the contents of sorbitol, xylitol, sucralose, and sodium benzoate in oral solutions have been included in the pharmacopoeias and patent literatures of various countries in the world. Therefore, for oral solutions, an accurate and effective detection and analysis method needs to be established to objectively reflect the dosages of sorbitol, xylitol, sucralose, and sodium benzoate in oral solutions, and provide a basis for the quality control and scientific and safe use of oral solutions. Summary of the Invention

[0005] The object of the present invention is to provide a method for simultaneously determining the contents of sorbitol, xylitol, sucralose, and sodium benzoate in a solution, controlling the dosages of the four substances; providing indicators for the research and development of oral solutions, and serving as an indicator for confirming the quality of drugs after the investigation of product stability and some changes (such as changes in formulation, production process, production site, and scale-up of production process). This method has simple operation, fast speed, strong specificity, and high accuracy.

[0006] The technical solution of the present invention is: using the solution after centrifugation of the solution as the test solution, then determining the contents of sorbitol, xylitol, sucralose, and sodium benzoate in the test solution, and then comparing with the labeled amounts of sorbitol, xylitol, sucralose, and sodium benzoate in the said solution.

[0007] The preparation method of the reference solution in the present invention is: accurately weigh appropriate amounts of sorbitol, xylitol, sucralose, and sodium benzoate reference substances and place them in a volumetric flask, dissolve with water and dilute to the scale, shake well, and use as the reference solution.

[0008] The present invention uses high performance liquid chromatography-tandem mass spectrometry to determine the contents of sorbitol, xylitol, sucralose, and sodium benzoate in the test solution.

[0009] The method of the present invention can be used to determine oral solutions, and can also be used to directly test the contents of sorbitol, xylitol, sucralose, and sodium benzoate in the test solution; preferably used for atomoxetine hydrochloride oral solution and the test solution.

[0010] The specific steps are as follows:

[0011] (1) Prepare the test solution: accurately measure an appropriate amount of the oral solution and place it in a volumetric flask, dilute to the scale with water, shake well, perform centrifugation treatment, and take the supernatant as the test solution;

[0012] (2) Preparation of reference solution: Weigh appropriate amounts of sorbitol, xylitol, sucralose, and sodium benzoate reference substances accurately and place them in a volumetric flask. Dissolve with water and dilute to the mark to prepare a solution containing approximately 40 μg of sorbitol, 300 μg of xylitol, 10 μg of sucralose, and 0.8 μg of sodium benzoate per 1 ml. Shake well, perform centrifugation, and take the supernatant as the reference solution;

[0013] (3) Determination of content: Take 10 μl each of the reference solution and each test solution, inject them into a high performance liquid chromatography - mass spectrometer respectively, and record the total ion current chromatogram; Calculate according to the external standard method with the peak area to calculate the contents of sorbitol, xylitol, sucralose, and sodium benzoate respectively.

[0014] Furthermore, the high performance liquid chromatography conditions adopted in the present invention are as follows: The chromatographic column is an octadecylsilane chemically bonded silica column. The acetonitrile - water - formic acid solution with a volume ratio of 100:900:1 is used as mobile phase A, and the acetonitrile - formic acid solution with a volume ratio of 1000:1 is used as mobile phase B. The column temperature of the chromatographic column is 40 °C, and the flow rate is 0.4 ml / min.

[0015] Furthermore, the conditions for gradient elution are as follows:

[0016]

[0017] Furthermore, the mass spectrometry conditions adopted in the present invention are as follows: Positive ion ionization mode (ESI + ) detection mode, MS scanning range: m / z 100 - 1000, spray voltage: 4.0 kV, nebulizer flow rate: 11 mL·min -1 , desolvation gas temperature: 350 °C, collision energy: 120 V, ion source gas pressure: 310 kPa.

[0018] Furthermore, the chromatographic column selected in the present invention is an Agilent Extend C18 chromatographic column.

[0019] The high performance liquid chromatography - mass spectrometry tandem method adopted in the present invention realizes an extremely short inspection time (running time) and can quickly and accurately quantitatively detect the contents of sorbitol, xylitol, sucralose, and sodium benzoate.

[0020] The beneficial effects of the present invention are as follows: The present invention provides a method for simultaneously and rapidly determining the contents of sorbitol, xylitol, sucralose, and sodium benzoate. A chromatographic column filled with octadecylsilane chemically bonded phase is used, and the high performance liquid chromatography - mass spectrometry tandem method is adopted. Acetonitrile, formic acid, and aqueous solution are used as mobile phases to detect and control the limits of the contents of sorbitol, xylitol, sucralose, and sodium benzoate in oral solution. This method has simple operation, is fast, has strong specificity, and high accuracy, and is of extremely important significance for realizing the quality control and safety assurance of oral solution.

[0021] The technical solutions of the present invention will be described in detail below through examples and drawings to better understand the advantages of the present invention. The provided examples should be construed as illustrative of the method of the present invention and should not limit the technical solutions and protection scope disclosed by the present invention in any way. Description of the Drawings

[0022] Figures 1 to 2 Total ion current chromatogram of blank solvent, characteristic ion chromatograms of sorbitol, xylitol, sucralose and sodium benzoate;

[0023] Figures 3 to 13 Total ion current chromatogram of reference substance solution, characteristic ion chromatograms of sorbitol, xylitol, sucralose and sodium benzoate;

[0024] Figures 14 to 20 Total ion current chromatogram of test solution, characteristic ion chromatograms of sorbitol, xylitol, sucralose and sodium benzoate. Detailed Description of the Embodiments

[0025] In the following detailed description of the embodiments, the reference substances, samples, reagents and instruments used are as follows:

[0026] Instrument: Agilent 1290-6540 high performance liquid chromatography-mass spectrometry tandem instrument;

[0027] Reagents: acetonitrile (mass spectrometry grade), formic acid (mass spectrometry grade), ultrapure water.

[0028] Chromatographic column: Agilent Extend C18 4.6×50mm, 1.8μm

[0029] Unless otherwise specified, the solvent mentioned in the present invention is ultrapure water.

[0030] Sorbitol, xylitol and sucralose are all commercially available, and the manufacturers are all Aladdin. The batch numbers are: H1921071 (sorbitol), H1711014 (xylitol), B1828061 (sucralose), and the contents are: 99.5% (sorbitol), 100% (xylitol), 98% (sucralose).

[0031] Sodium benzoate is purchased from the National Institutes for Food and Drug Control, batch number: 100433-201702, content 99.7%.

[0032] Atomoxetine hydrochloride oral solution is commercially available, the manufacturer is Eli Lilly Germany, batch number: 8048C; main component: atomoxetine hydrochloride, specification: 4mg / mL (calculated as C 17 H 21 NO).

[0033] Example 1 Specificity Test

[0034] Blank solvent: ultrapure water.

[0035] Stock solution of sodium benzoate: Accurately weigh 8 mg of sodium benzoate reference substance, place it in a 10-ml volumetric flask, dissolve it with water and dilute to the mark, shake well, and you will get it.

[0036] Reference substance solution: Accurately weigh 40 mg of sorbitol reference substance, 300 mg of xylitol reference substance, and 10 mg of sucralose reference substance, place them in a 100-ml volumetric flask. After adding 1 ml of the stock solution of sodium benzoate, dissolve it with water and dilute to the mark, shake well to obtain the stock solution of the reference substance; accurately measure 1 ml of the stock solution and place it in a 10-ml volumetric flask, dilute it to the mark with the solvent, shake well, and you will get it.

[0037] Using the chromatographic and mass spectrometric conditions of this invention patent, detect the blank solvent and the reference substance solution respectively, record the total ion current chromatogram, which are respectively Figures 1 to 13 as shown.

[0038] Result: No interfering peaks were detected at the corresponding positions of the ion absorption peaks of sorbitol, xylitol, sucralose, and sodium benzoate in the blank solvent solution, indicating that this method has good specificity for the determination of the contents of sorbitol, xylitol, sucralose, and sodium benzoate.

[0039] Example 2 System suitability test

[0040] Take 10 μl of the reference substance solution in Example 1, inject it into the high-performance liquid chromatography-mass spectrometer, and detect it according to the chromatographic and mass spectrometric conditions of this invention patent, and record the total ion current chromatogram. The test results are shown in Figures 3 to 14 and Table 1.

[0041] Table 1 Results of system suitability test

[0042]

[0043] In the total ion chromatogram of the reference substance solution, the RSD of the peak areas of each peak for 6 consecutive injections is less than 20.0%, meeting the requirements of system suitability.

[0044] Example 3 Determination test of the contents of sorbitol, xylitol, sucralose, and sodium benzoate in oral solution

[0045] Test solution: Accurately measure 1 ml of the sample and place it in a 100-ml volumetric flask, dilute it to the mark with water to obtain the stock solution; accurately measure 1 ml of the stock solution and place it in a 10-ml volumetric flask, dilute it to the mark with the solvent, shake well, and you will get it.

[0046] Take 10 μl of each of the test solution and the reference substance solution in Example 1, inject them into the high-performance liquid chromatography-mass spectrometer, and detect them according to the chromatographic and mass spectrometric conditions of this invention patent, and record the total ion current chromatogram.

[0047] According to the quantitative ions (m / z) of sorbitol, xylitol, sucralose and sodium benzoate in Table 1, extract each quantitative ion (m / z) from the total ion current chromatogram collected and perform integration, measure the peak area, and calculate the contents of sorbitol, xylitol, sucralose and sodium benzoate in the sample. The test results are shown in Figures 3 to 20 and Table 2.

[0048] Calculation formula:

[0049]

[0050] C sample: Concentration of the sample

[0051] A sample: Peak area of the sample

[0052] C reference: Concentration of the reference

[0053] A reference: Average peak area of the reference injected continuously for 6 times

[0054] Table 2 Results of the determination of the contents of sorbitol, xylitol, sucralose and sodium benzoate

[0055]

[0056] Note: [1] The original research quality standard of sodium benzoate stipulates that the content of sodium benzoate is 0.8 ± 0.08 mg / ml

[0057] [2] The sorbitol solution contains about 70% sorbitol. It can be calculated that the sorbitol content in the patented public prescription is 32.97 mg / ml (47.1 * 70%).

[0058] The measured contents of sorbitol, xylitol, sucralose and sodium benzoate are basically consistent with the prescription of the comparative patent.

Claims

1. A method for simultaneously determining the contents of sorbitol, xylitol, sucralose and sodium benzoate in a solution preparation, comprising the following steps: Take an appropriate amount of the solution and dilute it with water. Take the supernatant after centrifugation as the test solution, then determine the contents of sorbitol, xylitol, sucralose and sodium benzoate in the test solution, and then compare them with the labeled amounts of sorbitol, xylitol, sucralose and sodium benzoate in the solution preparation.

2. The method for simultaneously determining the contents of sorbitol, xylitol, sucralose and sodium benzoate in a solution preparation according to claim 1 or 2, characterized in that the method is as follows: (1) Prepare the test solution: Accurately measure 1 ml of the solution and dilute it to 100 ml with water to obtain a stock solution; accurately measure 1 ml of the stock solution and dilute it to 10 ml with water, shake well, and use it as the test solution; (2) Prepare the sodium benzoate stock solution: Accurately weigh the sodium benzoate reference substance, dissolve it with water and dilute it, shake well, and prepare a sodium benzoate stock solution with a sodium benzoate concentration of 0.8 mg / ml; (3) Prepare the stock solution of the reference solution: Accurately weigh the sorbitol reference substance, xylitol reference substance, and sucralose reference substance, accurately measure the sodium benzoate stock solution, dissolve it with water and dilute it, shake well, and prepare a stock solution of the reference solution containing 0.4 mg of sorbitol, 3 mg of xylitol, 0.1 mg of sucralose, and 0.004 mg of sodium benzoate per 1 ml; Accurately measure the stock solution of the reference solution, dilute it with water, shake well, and prepare a reference solution containing 40 μg of sorbitol, 300 μg of xylitol, 10 μg of sucralose, and 0.8 μg of sodium benzoate per 1 ml; (4) Determine the content: Use high performance liquid chromatography-tandem mass spectrometry to detect the reference solution and each test solution; calculate according to the external standard method with the peak area, and calculate the contents of sorbitol, xylitol, sucralose and sodium benzoate respectively.

3. The method according to claim 1 or 2, characterized in that the solution preparation is atomoxetine hydrochloride oral solution.

4. The method according to claim 1 or 2, characterized in that the solution preparation is the test solution.

5. The method according to claim 3, characterized in that The high performance liquid chromatography conditions were as follows: the chromatographic column was an octadecylsilyl silica gel column, the acetonitrile-water-formic acid solution with a volume ratio of 100:900:1 was used as mobile phase A, the acetonitrile-formic acid solution with a volume ratio of 1000:1 was used as mobile phase B, and gradient elution was performed; the column temperature of the chromatographic column was 40 °C and the flow rate was 0.4 ml / min; the mass spectrometry conditions were as follows: positive ion ionization mode (ESI + ) detection mode, MS scanning range: m / z 100-1000, spray voltage: 4.0 kV, nebulizer flow rate: 11 mL·min -1 , desolvation gas temperature: 350 °C, collision energy: 120 V, ion source gas pressure: 310 kPa.

6. The method according to claim 5, characterized in that the chromatographic column used in the selected high performance liquid chromatography method is Agilent Extend C18.

7. The method according to claim 4, characterized in that The high performance liquid chromatography conditions are as follows: the chromatographic column is an octadecylsilane-bonded silica gel column, the acetonitrile-water-formic acid solution with a volume ratio of 100:900:1 is used as mobile phase A, the acetonitrile-formic acid solution with a volume ratio of 1000:1 is used as mobile phase B, and gradient elution is performed; the column temperature of the chromatographic column is 40 °C and the flow rate is 0.4 ml / min; the mass spectrometry conditions are as follows: positive ion ionization mode (ESI + ) detection mode, MS scanning range: m / z 100-1000, spray voltage: 4.0 kV, nebulizer flow rate: 11 mL·min -1 , desolvation gas temperature: 350 °C, collision energy: 120 V, ion source gas pressure: 310 kPa.

8. The method according to claim 7, characterized in that the chromatographic column used in the selected high performance liquid chromatography method is Agilent Extend C18.

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