Method for determining content of N-nitroso tamsulosin in tamsulosin hydrochloride and application
The method of detecting N-nitrosotamsulosin in tamsulosin hydrochloride by LC-MS/MS solves the problem of ineffective detection in existing technologies, achieving rapid and accurate detection results and ensuring drug safety.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-26
- Publication Date
- 2026-04-07
AI Technical Summary
The lack of effective methods in the current technology for detecting the content of N-nitrosotamsulosin in tamsulosin hydrochloride makes it impossible to effectively control its residue in the drug, which poses a safety hazard.
The detection was performed using liquid chromatography-mass spectrometry (LC-MS/MS). An Agilent Infinity Lab Poroshell 120 EC-C18 column and an electrospray ionization (ESI) source were used. With specific chromatographic conditions and mass spectrometry parameters, the target impurities and matrix interferences were separated through a gradient elution program to achieve accurate quantification of N-nitrosotamsulosin.
This method enables rapid and accurate detection of N-nitrosotamsulosin in tamsulosin hydrochloride. It features short and precise detection time, high sensitivity, recovery rate of 90-111%, and linear range of 1.3-26.8 ng/ml, filling a detection gap and ensuring drug quality.
Smart Images
Figure CN121805448A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The application belongs to the technical field of N-nitroso tamsulosin content detection, and particularly relates to a determination method and application of N-nitroso tamsulosin content in tamsulosin hydrochloride. BACKGROUND
[0002] N-nitroso tamsulosin is an organic compound with a chemical formula of C 20 H 27 N3O6S, and a structure as shown in the following formula, which is a white solid powder, is a nitration product of small molecule drug tamsulosin hydrochloride, contains a warning structure, is a nitrosamine impurity, and has a nitroso group which can directly attack DNA to cause base alkylation, chain rupture or cross-linking, induce gene mutation and chromosome aberration, and has cumulative damage, so that the content thereof needs to be controlled at an acceptable level, and currently there is no report on the detection method of N-nitroso tamsulosin on the market.
[0003]
[0004] N-nitroso tamsulosin structure SUMMARY The application aims at the deficiencies of the prior art, and provides a determination method and application of N-nitroso tamsulosin content in tamsulosin hydrochloride. The determination method is short in time consumption and low in operation difficulty.
[0005] In a first aspect, the determination method of N-nitroso tamsulosin content in tamsulosin hydrochloride comprises the following steps: detecting a control sample solution by an LC-MS / MS method to obtain a standard curve; and detecting a test sample solution by the LC-MS / MS method to calculate the N-nitroso tamsulosin content in the test sample solution through the standard curve.
[0006] Further, the chromatographic conditions of the LC-MS / MS method comprise: using an Agilent Infinity Lab Poroshell120 EC-C18 chromatographic column with a size of 50 mm*2.1 mm*1.9 μm; the chromatographic column temperature is 35 DEG C, the sample injection disc temperature is not controlled, and the sample injection disc temperature comprises a sample injection port temperature; the sample injection amount is 2 μl.
[0007] Further, the mass spectrometry conditions of the LC-MS / MS method comprise: using a mass spectrometer as a detector and an ESI as an ion source; selecting an MRM ion collection mode, collecting ion pairs in a positive ion mode as 436 / 199 and 436 / 137, and collecting a quantitative ion pair as 436 / 199.
[0008] Further, the chromatographic conditions of the LC-MS / MS method comprise: a flow rate of 0.4 ml / min; a mobile phase A of 0.1% formic acid aqueous solution; and a mobile phase B of acetonitrile.
[0009] Further, the chromatographic conditions of the LC-MS / MS method comprise that the gradient elution program is:
[0011] Further, the preparation method of the control solution comprises that N-nitroso tamsulosin control sample is taken, diluent is added, and the control solution is prepared. Preferably, the diluent comprises acetonitrile; and the concentration of the diluent is 10-30%.
[0012] Further, the preparation method of the test solution comprises that tamsulosin hydrochloride raw material is taken, solvent is added, and the test solution is prepared.
[0013] Preferably, the solvent comprises acetonitrile; and the concentration of the solvent is 10-30%.
[0014] In the second aspect, according to the application of the determination method of the content of N-nitroso tamsulosin in tamsulosin hydrochloride, the determination method is applied to detect the residual amount of N-nitroso tamsulosin in tamsulosin hydrochloride.
[0015] Compared with the prior art, the beneficial effects of the present application are reflected in: (1) The detection time of the present application is short, and one sample can be run in 6 minutes, and the operation difficulty is low.
[0016] (2) The determination method of the present application can realize accurate quantification to 25ppm, and the sensitivity, accuracy and precision are better.
[0017] (3) The linear range of the determination method of the present application is 1.3-26.8ng / ml, and the linear relationship is good.
[0018] (4) The recovery rate of the present application can reach 90-111%, and the accuracy of the determination method of the present application is better.
[0019] (5) The present application fills the blank of the detection method of N-nitroso tamsulosin in tamsulosin hydrochloride in the current market, provides a reliable analysis means for drug quality control, and helps to ensure the safety of medication.
[0020] (6) The present method adopts gradient elution program, combines specific chromatographic column and mass spectrometry parameters, can effectively separate target impurities and matrix interference, and improves the selectivity and accuracy of detection. BRIEF DESCRIPTION OF DRAWINGS
[0021] The accompanying drawings are used to provide further understanding of the present application, and constitute a part of the specification, and are used to explain the present application together with embodiments of the present application, and do not constitute a limitation on the present application.
[0022] In the drawings: Figure 1 is a standard curve graph of the N-nitroso tamsulosin reference solution of the present application; Figure 2 is a spectrum graph of the blank solution of the present application; Figure 3 is a spectrum graph of the test solution of the present application; Figure 4 is a spectrum graph of the reference solution of the present application; Figure 5 is a spectrum graph of the 100% test solution of the present application. DETAILED DESCRIPTION
[0023] The present application will be described in detail below with reference to Examples, which are merely representative and not intended to limit the scope of the present application in any way. In addition, all reagents used in the following Examples are commercially available or can be synthesized according to the existing literature or known methods, and the reaction or test conditions not listed are also conventional technical means readily available to those skilled in the art, if not otherwise specified. The terms used in the present application are common in the art and can be clearly understood by those skilled in the art, and will not be described here.
[0024] It is particularly noted that, chromatographic column, Agilent Infinity Lab Poroshell 120 EC-C18, Agilent.
[0025] Method 1 The present application provides a method for determining the content of N-nitroso tamsulosin in tamsulosin hydrochloride, comprising the following steps: (1) Take an appropriate amount of N-nitroso tamsulosin reference substance, dilute it with 20% acetonitrile aqueous solution to prepare an appropriate concentration of N-nitroso tamsulosin reference solution as the reference solution; take tamsulosin hydrochloride raw material, dissolve it with 20% acetonitrile aqueous solution to prepare an appropriate concentration of test solution as the test solution; (2) Prepare a series of N-nitroso tamsulosin reference solutions, detect them by LC-MS / MS method, respectively obtain the peak area of N-nitroso tamsulosin in the reference solution, and perform standard curve fitting with the solution concentration to obtain a standard curve; (3) Take the test solution and detect it by LC-MS / MS method to obtain the peak area of N-nitroso tamsulosin in the test solution, and calculate the content of N-nitroso tamsulosin in the test solution by external standard curve method.
[0026] In the above steps (2) and (3), the conditions of LC-MS / MS method are as follows: Agilent Infinity Lab Poroshell 120 EC-C18 column, 50 mm x 2.1 mm x 1.9 μm, column temperature 35 ℃, injection volume 2 μl, uncontrolled temperature of injection tray; detector is mass spectrometer, ion source is ESI; selected ion acquisition mode (MRM), positive ion mode acquisition ion pair is 436 / 199 and 436 / 137, quantitative ion pair is 436 / 199; flow rate is 0.4 ml / min; mobile phase A is 0.1% formic acid aqueous solution, mobile phase B is acetonitrile, gradient elution program is as follows Table 1: Table 1
[0027] Referring to Method 1, the specific implementation process of Method 1 is given as follows: Example 1 The determination method of N-nitroso tamsulosin content in tamsulosin hydrochloride is as follows: Table 2 Reference solution
[0028] Table 3
[0029] (1) Referring to Table 2, N-nitroso tamsulosin reference substance is taken, 20% acetonitrile aqueous solution is used as diluent, and N-nitroso tamsulosin reference solution of different concentrations (including L-1~L-5) is prepared as reference solution; Referring to Table 3, tamsulosin hydrochloride raw material is taken, 20% acetonitrile aqueous solution is used as solvent, and test solution of different concentrations is prepared as test solution; (2) The series of N-nitroso tamsulosin reference solution prepared in step (1) is detected by LC-MS / MS method, the peak area of N-nitroso tamsulosin in the reference solution is obtained, and the standard curve is fitted with the solution concentration to obtain the standard curve; the standard curve of reference solution is referred to Figure 1 and Table 2.
[0030] (3) The test solution prepared in step (1) is detected by LC-MS / MS method, the peak area of N-nitroso tamsulosin in the test solution is obtained, and the content of N-nitroso tamsulosin in the test solution is calculated by external standard curve method. The results are referred to Table 3.
[0031] In addition, SPL-1 spectrum can be referred to Figure 3 . Figure 2 and Figure 4 are both specificity related spectra.
[0032] Example 2 In Example 1, the conditions of the LC-MS / MS method in Step (2) and Step (3) are as follows: An Agilent Infinity Lab Poroshell 120 EC-C18 chromatographic column, 50 mm x 2.1 mm x 1.9 μm, was used, the column temperature was 35°C, the injection volume was 2 μl, and the injection tray was not temperature-controlled; the detector was a mass spectrometer, the ion source was ESI; the ion selection acquisition mode (MRM) was used, the positive ion mode was used to collect ion pairs 436 / 199 and 436 / 137, the quantitative ion pair was 436 / 199; the flow rate was 0.4 ml / min; mobile phase A was 0.1% formic acid aqueous solution, mobile phase B was acetonitrile, and the gradient elution program is shown in Table 1.
[0033] Example 3 In this example, compared with Example 1 and Example 2, the accuracy of the determination method of the application was further tested, and the steps were as follows: Referring to Table 3, in 9 samples of tamsulosin hydrochloride not containing N-nitroso-tamsulosin (10%-1, 10%-2, 10%-3, 100%-1, 100%-2, 100%-3, 150%-1, 150%-2, 150%-3), different levels of N-nitroso-tamsulosin were added (i.e. the amount added), the content of N-nitroso-tamsulosin after addition was determined (i.e. the amount measured), and the recovery rate was calculated.
[0034] The recovery rate formula is: recovery rate = (measured amount - original amount) / added amount.
[0035] Result analysis: according to the method accuracy regulation, the recovery rate should be within the range of 80%-120%, and the application meets this regulation.
[0036] Example 4 In this example, compared with Examples 1-3, the repeatability of the determination method of the application was further tested, and the steps were as follows: Table 4
[0037] Referring to Table 4, in 6 samples of tamsulosin hydrochloride not containing N-nitroso-tamsulosin (100%-1, 100%-2, 100%-3, 100%-4, 100%-5, 100%-6), the same level of N-nitroso-tamsulosin was added (i.e. the amount added), the content of N-nitroso-tamsulosin after addition was determined, and the recovery rate was calculated. The 100%-1 spectrum can also be referred to Figure 5 .
[0038] Application: Generally, qualified tamsulosin hydrochloride bulk drug does not contain N-nitroso tamsulosin, if the bulk drug degrades, the determination method of the application can be used to detect whether N-nitroso tamsulosin is contained in the tamsulosin hydrochloride bulk drug, i.e. the residual amount, so as to monitor the quality of the tamsulosin hydrochloride bulk drug.
[0039] The above examples are only used to illustrate the technical solutions of the present application, but not limit the same, and the purpose is to enable the person skilled in the art to understand the content of the present application and to implement it, any equivalent changes or modifications made according to the spirit and essence of the present application should be covered within the protection scope of the present application.
Claims
1. A method for determining the N-nitrosotamsulosin content in tamsulosin hydrochloride, characterized in that, The procedure includes the following steps: detecting the reference solution using LC-MS / MS to obtain a standard curve; detecting the test solution using LC-MS / MS and calculating the N-nitrosotamsulosin content in the test solution using the standard curve.
2. The method for determining the N-nitrosotamsulosin content in tamsulosin hydrochloride according to claim 1, characterized in that, The chromatographic conditions for LC-MS / MS included: an Agilent Infinity Lab Poroshell 120 EC-C18 column (50 mm × 2.1 mm × 1.9 μm); a column temperature of 35 °C; no control of the injection tray temperature (including the injection port temperature); and an injection volume of 2 μl.
3. The method for determining the N-nitrosotamsulosin content in tamsulosin hydrochloride according to claim 1, characterized in that, The mass spectrometry conditions for LC-MS / MS include: a mass spectrometer detector and an ESI ion source; selection of MRM ion acquisition mode, with positive ion mode acquiring ion pairs of 436 / 199 and 436 / 137, and quantitative ion pair of 436 / 199.
4. The method for determining the N-nitrosotamsulosin content in tamsulosin hydrochloride according to claim 1, characterized in that, The chromatographic conditions for LC-MS / MS included: a flow rate of 0.4 ml / min; mobile phase A was 0.1% formic acid aqueous solution, and mobile phase B was acetonitrile.
5. The method for determining the N-nitrosotamsulosin content in tamsulosin hydrochloride according to claim 1, characterized in that, The chromatographic conditions for the LC-MS / MS method include: a gradient elution program as follows: 。 6. The method for determining the N-nitrosotamsulosin content in tamsulosin hydrochloride according to claim 1, characterized in that, The preparation method of the reference solution includes: taking N-nitrosotamsulosin reference standard, adding diluent, and preparing the reference solution.
7. The method for determining the N-nitrosotamsulosin content in tamsulosin hydrochloride according to claim 6, characterized in that, The diluent includes acetonitrile; the concentration of the diluent is 10-30%.
8. The method for determining the N-nitrosotamsulosin content in tamsulosin hydrochloride according to claim 1, characterized in that, The preparation method of the test solution includes: taking tamsulosin hydrochloride raw material, adding solvent, and preparing the test solution.
9. The method for determining the N-nitrosotamsulosin content in tamsulosin hydrochloride according to claim 8, characterized in that, The solvent includes acetonitrile; the concentration of the solvent is 10-30%.
10. The application of the method for determining the N-nitrosotamsulosin content in tamsulosin hydrochloride according to any one of claims 1-9, characterized in that, This assay method is used to detect the residual amount of N-nitrosotamsulosin in tamsulosin hydrochloride.