Method for determining content of sodium chloride in tirofiban hydrochloride sodium chloride injection

Abstract

The invention relates to the field of chemical detection and the technical field of pharmaceutical analysis, and discloses a method for determining the content of sodium chloride in tirofiban hydrochloride sodium chloride injection, which comprises the following steps: taking a sodium chloride reference substance, and preparing a reference substance solution by using a phosphoric acid solution; taking the tirofiban hydrochloride sodium chloride injection, preparing a test solution by using a phosphoric acid solution, respectively and precisely measuring the reference solution and the test solution, carrying out liquid chromatography detection, recording a chromatogram, and calculating the content of sodium chloride in the tirofiban hydrochloride sodium chloride injection by peak area according to an external standard method. According to the method for determining the content of sodium chloride in the tirofiban hydrochloride sodium chloride injection, the content of sodium chloride can be accurately determined, sodium chloride can be separated from other substances, accurate detection of sodium chloride is achieved, and therefore the product quality of the tirofiban hydrochloride sodium chloride injection is better controlled. And the mobile phase is used as a test solvent, so that the operation steps are simplified, the detection data is accurate, the stability is good, the safety is high, and the detection cost is low.

Description

Technical Field

[0001] The invention relates to the fields of chemical detection and drug analysis technology, and in particular to a method for determining the sodium chloride content in tirofiban hydrochloride sodium chloride injection. Background Art

[0002] Tirofiban hydrochloride sodium chloride injection is a non-peptide platelet surface glycoprotein GPⅡb / Ⅲa receptor antagonist used to treat acute coronary syndrome. Since its launch, its clinical application value has been widely recognized. The formula of this injection contains sodium chloride, which, as an electrolyte balance regulator, plays an important role in maintaining electrolyte balance in the body. However, according to the general rules of the four parts of the 2020 edition of the "Chinese Pharmacopoeia", the current method for determining the sodium chloride content adopts titration, using ammonium ferric sulfate as an indicator and ammonium thiocyanate as a titrant. This method has certain human errors, and because the content of sodium chloride in the injection is relatively small, and the titration method requires a larger amount of sodium chloride, it is not suitable for the accurate determination of lower concentrations of sodium chloride in the injection. Summary of the invention

[0003] In view of the above problems existing in the prior art, the present application provides a method for determining the sodium chloride content in tirofiban hydrochloride sodium chloride injection, so as to improve the accuracy and stability of the determination of the sodium chloride content in the injection.

[0004] To solve the above problems, the present invention provides the following technical solutions:

[0005] The present application provides a method for determining the sodium chloride content in tirofiban hydrochloride sodium chloride injection, comprising the following steps:

[0006] Take the sodium chloride reference substance and prepare a reference solution with a solvent;

[0007] Take tirofiban hydrochloride sodium chloride injection and prepare the test solution with solvent:

[0008] Accurately measure the reference solution and the test solution for liquid chromatography detection, record the chromatogram, and calculate the content of sodium chloride in tirofiban hydrochloride sodium chloride injection by peak area according to the external standard method;

[0009] The conditions for liquid chromatography detection are:

[0010] The model of the chromatographic column is Shodex SUGAR SH1011;

[0011] The specifications of the chromatographic column are 8.0 mm × 300 mm × 6 μm;

[0012] The column temperature is 45-55°C;

[0013] The mobile phase was phosphoric acid solution;

[0014] Flow rate 0.5-0.7 mL / min;

[0015] The injection volume of the reference solution and the test solution was 10 μL respectively;

[0016] The detection wavelength is 180-200nm.

[0017] In one embodiment of the present application, the solvent is a phosphoric acid solution.

[0018] In one embodiment of the present application, the concentration of the reference solution is 0.18 mg / mL.

[0019] In one embodiment of the present application, the concentration of the test solution is 0.001 mg / mL.

[0020] In one embodiment of the present application, the concentration of the phosphoric acid solution is 4.5 mg / mL.

[0021] In one embodiment of the present application, the column temperature of the chromatographic column is 50°C.

[0022] In one embodiment of the present application, the flow rate is 0.6 mL / min.

[0023] In one embodiment of the present application, the detection wavelength is 190 nm.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] (1) The method for determining the sodium chloride content in tirofiban hydrochloride sodium chloride injection provided by the present invention can accurately detect the sodium chloride in tirofiban hydrochloride sodium chloride injection, separate the sodium chloride from other substances in the injection, improve the accuracy and repeatability of the sodium chloride content determination, and thus better control the product quality of tirofiban hydrochloride sodium chloride injection;

[0026] (2) The method for determining the sodium chloride content in tirofiban hydrochloride sodium chloride injection provided by the present invention uses the mobile phase as the solvent, simplifies the operation steps, and has accurate test data, good stability, high safety, and low test cost;

[0027] (3) The method for determining the sodium chloride content in tirofiban hydrochloride sodium chloride injection provided by the present invention has a good linear relationship for sodium chloride, and the linear correlation coefficient can reach above 0.999. The determination method of the present invention has good quantitative analysis capability. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 is a chromatogram of tirofiban hydrochloride solution;

[0029] Figure 2 is the chromatogram of blank excipient;

[0030] Figure 3 is the chromatogram of the reference solution;

[0031] Figure 4 is the chromatogram of the test solution;

[0032] Figure 5 is the chromatogram of linear solution 1 (80% concentration level);

[0033] Figure 6 is the chromatogram of linear solution 2 (90% concentration level);

[0034] Figure 7 is the chromatogram of linear solution 3 (100% concentration level);

[0035] Figure 8 is the chromatogram of linear solution 4 (110% concentration level);

[0036] Fig. 9 is the chromatogram of linear solution 5 (120% concentration level);

[0037] Fig.10 This is the chromatogram of the accuracy solution (80% low concentration);

[0038] Fig.11 is the chromatogram of the accuracy solution (100% medium concentration);

[0039] Fig.12 This is the chromatogram of the accuracy solution (120% high concentration). DETAILED DESCRIPTION

[0040] The following will provide a clear and complete description of the technical solutions in the embodiments of the present application in conjunction with the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all of the embodiments.

[0041] The "range" disclosed in this application is defined in the form of a lower limit and an upper limit, and a given range is defined by selecting a lower limit and an upper limit, and the selected lower limit and upper limit define the boundaries of the particular range. The range defined in this way can be inclusive or exclusive of the end values, and can be arbitrarily combined, that is, any lower limit can be combined with any upper limit to form a range.

[0042] Unless otherwise specified, when the present invention relates to the percentage between liquids, the percentage is volume / volume percentage; when the present invention relates to the percentage between liquids and solids, the percentage is volume / weight percentage; when the present invention relates to the percentage between solids and liquids, the percentage is weight / volume percentage; the rest are weight / weight percentages.

[0043] Example 1

[0044] The high performance liquid chromatography method was used for determination, and the operation steps were as follows:

[0045] (1) Establishment of chromatographic conditions and system suitability: A Shodex SUGAR SH1011 (8.0 mm × 300 mm × 6 μm) chromatographic column was used, with phosphoric acid solution (4.5 g of phosphoric acid added to 1000 mL of water) as the mobile phase, the detection wavelength was 190 nm, the flow rate was 0.6 mL / min, and the column temperature was 50°C;

[0046] (2) Preparation of reference solution: Take an appropriate amount of sodium chloride reference substance, weigh it accurately, add phosphoric acid solution and ultrasonically dissolve it and dilute it to make a solution containing about 0.18 mg per 1 mL;

[0047] (3) Preparation of test solution: Accurately measure 1 mL of tirofiban hydrochloride and sodium chloride injection, place it in a 50 mL volumetric flask, add phosphoric acid solution to dilute to the mark, shake well, and prepare a test solution with a concentration of 0.001 mg / mL;

[0048] (4) Determination: Accurately measure 10 μL of the reference solution and the test solution respectively and inject them into the liquid chromatograph. Record the chromatogram and calculate the peak area using the external standard method.

[0049] Example 2

[0050] The high performance liquid chromatography method was used for determination, and the operation steps were as follows:

[0051] (1) Establishment of chromatographic conditions and system suitability: A Shodex SUGAR SH1011 (8.0 mm × 300 mm × 6 μm) chromatographic column was used, with phosphoric acid solution (4.5 g of phosphoric acid added to 1000 mL of water) as the mobile phase, the detection wavelength was 190 nm, the flow rate was 0.5 mL / min, and the column temperature was 50°C;

[0052] (2) Preparation of reference solution: Take an appropriate amount of sodium chloride reference substance, weigh it accurately, add phosphoric acid solution and ultrasonically dissolve it and dilute it to make a solution containing about 0.18 mg per 1 mL;

[0053] (3) Preparation of test solution: Accurately measure 1 mL of tirofiban hydrochloride and sodium chloride injection, place it in a 50 mL volumetric flask, add phosphoric acid solution to dilute to the mark, shake well, and prepare a test solution with a concentration of 0.001 mg / mL;

[0054] (4) Determination: Accurately measure 10 μL of the reference solution and the test solution respectively and inject them into the liquid chromatograph. Record the chromatogram and calculate the peak area using the external standard method.

[0055] Example 3

[0056] The high performance liquid chromatography method was used for determination, and the operation steps were as follows:

[0057] (1) Establishment of chromatographic conditions and system suitability: A Shodex SUGAR SH1011 (8.0 mm × 300 mm × 6 μm) chromatographic column was used, with phosphoric acid solution (4.5 g of phosphoric acid added to 1000 mL of water) as the mobile phase, the detection wavelength was 190 nm, the flow rate was 0.7 mL / min, and the column temperature was 50°C;

[0058] (2) Preparation of reference solution: Take an appropriate amount of sodium chloride reference substance, weigh it accurately, add phosphoric acid solution and ultrasonically dissolve it and dilute it to make a solution containing about 0.18 mg per 1 mL;

[0059] (3) Preparation of test solution: Accurately measure 1 mL of tirofiban hydrochloride and sodium chloride injection, place it in a 50 mL volumetric flask, add phosphoric acid solution to dilute to the mark, shake well, and prepare a test solution with a concentration of 0.001 mg / mL;

[0060] (4) Determination: Accurately measure 10 μL of the reference solution and the test solution respectively and inject them into the liquid chromatograph. Record the chromatogram and calculate the peak area using the external standard method.

[0061] Example 4

[0062] The high performance liquid chromatography method was used for determination, and the operation steps were as follows:

[0063] (1) Establishment of chromatographic conditions and system suitability: A Shodex SUGAR SH1011 (8.0 mm × 300 mm × 6 μm) chromatographic column was used, with phosphoric acid solution (4.5 g of phosphoric acid added to 1000 mL of water) as the mobile phase, the detection wavelength was 190 nm, the flow rate was 0.6 mL / min, and the column temperature was 45°C;

[0064] (2) Preparation of reference solution: Take an appropriate amount of sodium chloride reference substance, weigh it accurately, add phosphoric acid solution and ultrasonically dissolve it and dilute it to make a solution containing about 0.18 mg per 1 mL;

[0065] (3) Preparation of test solution: Accurately measure 1 mL of tirofiban hydrochloride and sodium chloride injection, place it in a 50 mL volumetric flask, add phosphoric acid solution to dilute to the mark, shake well, and prepare a test solution with a concentration of 0.001 mg / mL;

[0066] (4) Determination: Accurately measure 10 μL of the reference solution and the test solution respectively and inject them into the liquid chromatograph. Record the chromatogram and calculate the peak area using the external standard method.

[0067] Example 5

[0068] The high performance liquid chromatography method was used for determination, and the operation steps were as follows:

[0069] (1) Establishment of chromatographic conditions and system suitability: A Shodex SUGAR SH1011 (8.0 mm × 300 mm × 6 μm) chromatographic column was used, with phosphoric acid solution (4.5 g of phosphoric acid added to 1000 mL of water) as the mobile phase, the detection wavelength was 190 nm, the flow rate was 0.6 mL / min, and the column temperature was 55°C;

[0070] (2) Preparation of reference solution: Take an appropriate amount of sodium chloride reference substance, weigh it accurately, add phosphoric acid solution and ultrasonically dissolve it and dilute it to make a solution containing about 0.18 mg per 1 mL;

[0071] (3) Preparation of test solution: Accurately measure 1 mL of tirofiban hydrochloride and sodium chloride injection, place it in a 50 mL volumetric flask, add phosphoric acid solution to dilute to the mark, shake well, and prepare a test solution with a concentration of 0.001 mg / mL;

[0072] (4) Determination: Accurately measure 10 μL of the reference solution and the test solution respectively and inject them into the liquid chromatograph. Record the chromatogram and calculate the peak area using the external standard method.

[0073] In order to further verify the feasibility of the present invention, a series of tests were carried out, as follows:

[0074] 1. Separation of sodium chloride from other substances

[0075] Weigh tirofiban hydrochloride and dilute it with a solvent to make a 1 μg / mL solution.

[0076] Weigh 54 mg of sodium citrate and 3.2 mg of anhydrous citric acid, add 100 mL of water to dissolve, measure 4 mL, place in a 200 mL volumetric flask, add solvent to dilute to the scale, shake well, and use as the blank excipient solution.

[0077] Test solution: Accurately measure 1.0 mL of the product, place it in a 50 mL volumetric flask, add solvent to dilute to the scale, and shake well.

[0078] Reference solution: Take about 18 mg of sodium chloride reference substance, weigh accurately, place in a 100 mL volumetric flask, add solvent to dissolve by ultrasonication and dilute to the scale, and shake well.

[0079] Take tirofiban hydrochloride solution, blank excipient solution, reference substance solution and test solution, and determine them by the above method. The HPLC spectrum is shown in Figure 1 to Figure 4 , the test results are shown in Table 1.

[0080] Table 1 Separation of sodium chloride from other substances

[0081] name Blank Accessories Tirofiban Hydrochloride Reference solution Test solution Retention time (min) No peak No peak 7.905 7.904

[0082] The results showed that the separation effect of sodium chloride and other substances in tirofiban hydrochloride sodium chloride injection was good, and other substances did not interfere with the detection of sodium chloride.

[0083] 2. Linearity of Sodium Chloride

[0084] Take sodium chloride reference substance and add mobile phase solution to obtain linear solutions with concentrations of 143.5 μg / mL, 161.5 μg / mL, 179.4 μg / mL, 197.4 μg / mL, and 215.3 μg / mL, respectively.

[0085] Take the above linear solution and detect it using the method of Example 1. The HPLC spectrum is shown in Figures 5 to 9 The test results are shown in Table 2.

[0086] Table 2 Linearity of sodium chloride

[0087]

[0088] The results show that the linear relationship of sodium chloride is good, and the linear correlation coefficient can reach above 0.999.

[0089] 3. Accuracy of Sodium Chloride

[0090] Reference substance stock solution: Accurately weigh an appropriate amount of sodium chloride, dissolve it in a solvent and dilute it to make a solution containing approximately 0.9 mg per 1 mL.

[0091] Accurately measure 4mL, 5mL, and 6mL of the reference stock solution, respectively, into a 25mL volumetric flask, and dilute to scale with blank excipients. Prepare three portions of each concentration in parallel to prepare an accuracy solution.

[0092] Take the above accuracy solution and use the method of Example 1 to detect it. The HPLC spectrum is shown in Figures 10-12 The test results are shown in Table 3.

[0093] Table 3 Accuracy of sodium chloride

[0094]

[0095] The results showed that the sodium chloride content was accurate.

[0096] The present application is described in detail above in conjunction with specific implementation methods and exemplary embodiments, but these descriptions cannot be understood as limiting the present application. Those skilled in the art understand that, without departing from the spirit and scope of the present application, the technical solution of the present application and its implementation methods may be subjected to a variety of equivalent substitutions, modifications or improvements, all of which fall within the scope of the present application.

Claims

1. A method for determining the sodium chloride content in tirofiban hydrochloride sodium chloride injection, characterized in that: The following steps are involved: Take the sodium chloride reference substance and prepare a reference solution with phosphoric acid solution; Take tirofiban hydrochloride sodium chloride injection and prepare the test solution with phosphoric acid solution: Accurately measure the reference solution and the test solution for liquid chromatography detection, record the chromatogram, and calculate the content of sodium chloride in tirofiban hydrochloride sodium chloride injection by peak area according to the external standard method; The conditions for the liquid chromatography detection are: The model of the chromatographic column is Shodex SUGAR SH1011; The specifications of the chromatographic column are 8.0 mm × 300 mm × 6 μm; The column temperature is 45-55°C; The mobile phase was phosphoric acid solution; Flow rate 0.5-0.7 mL / min; The injection volume of the reference solution and the test solution was 10 μL respectively; The detection wavelength is 180-200nm.

2. The method for determining the sodium chloride content in tirofiban hydrochloride sodium chloride injection according to claim 1, characterized in that: The concentration of the reference solution is 0.18 mg / mL.

3. The method for determining the sodium chloride content in tirofiban hydrochloride sodium chloride injection according to claim 1, characterized in that: The concentration of the test solution is 0.001 mg / mL.

4. The method for determining the sodium chloride content in tirofiban hydrochloride sodium chloride injection according to claim 1, characterized in that: The concentration of the phosphoric acid solution is 4.5 mg / mL.

5. The method for determining the sodium chloride content in tirofiban hydrochloride sodium chloride injection according to claim 1, characterized in that: The column temperature of the chromatographic column is 50°C.

6. The method for determining the sodium chloride content in tirofiban hydrochloride sodium chloride injection according to claim 1, characterized in that: The flow rate was 0.6 mL / min.

7. The method for determining the sodium chloride content in tirofiban hydrochloride sodium chloride injection according to claim 1, characterized in that: The detection wavelength is 190nm.

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