Method for determining contents of six elements in amine silver mesylate B powder injection preparation for injection

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a method for determining the contents of six elements in an amine silver mesylate lactone B powder injection preparation for injection. The method comprises the following steps: preparing a diluent, preparing a standard adding solution, preparing an element stock solution, preparing a standard curve solution, verifying linear parameters, verifying a limit of quantitation (LOQ), verifying specificity and verifying accuracy. Therefore, the content can be accurately measured, and clinical medication standards can be guided.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for determining the contents of six elements in an injection of silver amine mesylate lactone B powder injection preparation. Background Art

[0002] Percutaneous coronary intervention (PCI) is currently the main method for treating coronary heart disease, which has significantly reduced the incidence and mortality of cardiovascular events. However, the incidence of coronary artery thrombosis and vascular restenosis after this type of surgery is as high as 30% to 60%, which has become a serious obstacle to the further development of coronary heart disease treatment.

[0003] Based on the mechanism of silver amine lactone mesylate B in antagonizing PAF and its good efficacy in antiplatelet aggregation, it can be used to prevent thrombotic diseases. Percutaneous coronary intervention (PCI) significantly reduces the incidence and mortality of cardiovascular events, but the incidence of coronary artery restenosis after surgery is as high as 30% to 60%. This large postoperative population requires long-term or even lifelong oral antithrombotic drugs.

[0004] Currently, clopidogrel (Plavix) and aspirin are commonly used in clinical treatment of such diseases. Long-term use of the former can lead to drug resistance, resulting in loss of efficacy, the risk of myocardial infarction, and the serious adverse reaction of spontaneous bleeding; the latter has the adverse reaction of serious gastrointestinal mucosal damage, and patients with long-term medication have low tolerance. The development of silver amine lactone mesylate B for this indication has a very broad application prospect because of its high anticoagulant effect, no bleeding risk, good safety, and high tolerance. It can effectively prevent the formation of thrombus and restenosis of blood vessels after PCI treatment.

[0005] Inductively coupled plasma mass spectrometry (ICP-MS) is an inorganic multi-element analysis technique that uses inductively coupled plasma as an ion source and mass spectrometer for detection. The sample to be analyzed is usually introduced into an argon gas flow in the form of an aerosol of an aqueous solution, and then enters the central region of an argon plasma at atmospheric pressure excited by radio frequency energy. The high temperature of the plasma desolvates, vaporizes, dissociates, and ionizes the sample. Part of the plasma passes through different pressure zones and enters a vacuum system, where positive ions are pulled out and separated according to their mass-to-charge ratio. The detector converts the ions into electron pulses, which are then counted by an integral measurement circuit. The size of the electron pulse is related to the concentration of the analyzed ions in the sample. By comparing with known standards or reference materials, quantitative analysis of trace elements in unknown samples is achieved. Each element that occurs in nature has a simple or several isotopes, and the signal given by each specific isotope ion is linearly related to the concentration of the element in the sample.

[0006] The animal model of subacute thrombosis after PCI surgery has not yet been developed maturely in China. The present invention provides a method for determining the contents of six elements in an injection of silver amine lactone mesylate B powder injection preparation, so as to accurately measure the contents and guide clinical medication standards. Summary of the invention

[0007] The invention provides a method for determining the contents of six elements in an injection of silver amine mesylate lactone B powder injection preparation, so as to accurately measure the contents and guide clinical drug use standards.

[0008] To achieve the above object, the present invention provides the following technical solutions:

[0009] A method for determining the contents of six elements in a silver amine lactone mesylate B powder injection preparation for injection, characterized in that it comprises the following steps:

[0010] Step 1. Prepare diluent: Take 100 mL of nitric acid and dilute it to 1000 mL with water;

[0011] Step 2. Prepare the spike solution: Take the standard solutions of B, Si, and Sr elements and prepare them into a solution with a concentration of 100 μg / mL according to the table below;

[0012] Step 3. Prepare element stock solution: Take 100 μg / mL Al, B, Ba, Si, Sr, Ti element solution and diluent to prepare element stock solution;

[0013] Step 4. Prepare standard curve solution: Take B, Ba, Sr and Ti spike solutions and diluent to prepare standard curve solution;

[0014] Step 5. Take AI spike solution and diluent to prepare standard curve solution;

[0015] Step 6. Take Si spike solution and diluent to prepare standard curve solution;

[0016] Step 7. Prepare a specific solution: take 1.0 mL of pure water into a digestion tank, add 7 mL of nitric acid, pre-digest at 120°C for 20 min, seal after cooling, perform microwave digestion according to the table below, cool after digestion, transfer the contents to a 50 mL volumetric flask with pure water, and dilute to the mark;

[0017] Step 8. Prepare sample solution: Take 10 bottles of medicine, open them respectively and dissolve the contents, then pour them into the same plastic container and dilute to 50 ml as sample mother solution; take 1.0 ml of sample mother solution into a digestion tank, and prepare it in the same way as the "specific solution" starting from "add 7 ml of nitric acid"; prepare 2 portions in parallel using the same method;

[0018] Step 9. Prepare the accuracy solution: Low concentration level: Take 1.0 mL of the sample stock solution and place it in a digestion tank. Add 0.2 mL of the spike solution. Prepare the solution in the same way as the "specific solution" starting from "add 7 mL of nitric acid". The spike concentration is 8 μg / L. Prepare three replicates in parallel using the same method.

[0019] Step 10. Prepare medium-concentration solution: Take 1.0 mL of sample stock solution and place it in a digestion tank. Add 0.5 mL of spike solution. Prepare the solution in the same way as the "specific solution" starting from "add 7 mL of nitric acid". The spike concentration is 20 μg / L. Prepare three times in parallel using the same method.

[0020] Step 11. Prepare high-concentration level solution: Take 1.0 mL of sample mother solution and place it in a digestion tank. Add 0.75 mL of spike solution. Prepare the solution in the same way as the "specific solution" starting from "add 7 mL of nitric acid". The spike concentration is 30 g / L. Prepare three times in parallel using the same method.

[0021] The following steps are also included:

[0022] Step 12. Verify linear parameters: Take the standard curve solution for analysis, and the instrument automatically gives a linear equation. The correlation coefficient r of the linear equation of all elements should be no less than 0.995.

[0023] Step 13. Verify the limit of quantitation LOQ: Take a specific solution, inject 11 injections continuously for analysis, and calculate the SD of the test results of the 11 injections; prepare a standard solution with a limit of 10SD as the instrument LOQ solution, and calculate the recovery rate according to the linear equation. The recovery rate should be between 70% and 150%;

[0024] Step 14. Verify specificity: Take a sample of the specific solution for analysis. The concentration of all elements measured should not exceed the concentration of the instrument LOQ solution.

[0025] Step 15. Verify accuracy: Take the accuracy solution for analysis and calculate the recovery rate according to the linear equation. The recovery rate should be between 70% and 150%.

[0026] The calculation formula for the verification accuracy in step 15 is:

[0027] Sample LOQ (μg / g) = instrument LOQ (μg / L) × dilution factor × loading volume (Lg);

[0028] Sample result (μg / g) = instrument measured (μg / L) × dilution factor × loading volume (I / g);

[0029]

[0030] Added (μg) = spike concentration (μg / L) × added volume (L); Measured (μg) = accuracy result (μg / L) × solution volume (L);

[0031]

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] The invention provides a method for determining the contents of six elements in an injection of silver amine mesylate lactone B powder injection preparation, which has high measurement accuracy. DETAILED DESCRIPTION

[0034] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0035] This example is used to demonstrate the accuracy of using ICP-MS to determine the six elements (Al, B, Ba, Si, Sr and Ti) in the injection product XQ powder injection provided by Jiangsu Kefeiping Pharmaceutical Co., Ltd.

[0036] Table 1. XQ powder product sample information table

[0037]

[0038] Table 2. Instrument details

[0039]

[0040] Table 3. Reagent list

[0041]

[0042]

[0043] Table 4. Solution list

[0044]

[0045] Table 5. Internal standard solution list

[0046] name SGS ID 25ppb (In, Bi, Sc, Tb, Y) 21-90GSXI

[0047] 1. Verification process

[0048] 1.1 Solution preparation

[0049] 1.1.1 Diluent: Take 100 mL of nitric acid and dilute it to 1000 mL with water.

[0050] 1.1.2 Spiked solution: Take the standard solutions of B, Si and Sr elements and prepare them into a solution with a concentration of 100 μg / mL according to the table below.

[0051] Table 6. Dilution solution details

[0052]

[0053]

[0054] Take 100 μg / mL Al, B, Ba, Si, Sr, Ti element solutions and diluents, and prepare element stock solutions according to the table below.

[0055] Table 7. Element stock solution details

[0056]

[0057] 1.1.3 Standard curve solution: Take B, Ba, Sr and Ti spike solutions and diluents and prepare the standard curve solution according to the table below.

[0058] Table 8. Standard curve solution table

[0059]

[0060] Take AI spike solution and diluent and prepare standard curve solution according to the table below.

[0061] Table 9. Standard curve solution table after taking AI spike solution and diluent

[0062]

[0063]

[0064] Take Si spike solution and diluent and prepare standard curve solution according to the table below.

[0065] Table 10. Standard curve solution table after taking Si spike solution and diluent

[0066]

[0067] 1.1.4 Specific solution

[0068] Take 1.0mL of pure water in a digestion tank, add 7mL of nitric acid, pre-digest at 120℃ for 20min, seal after cooling, and perform microwave digestion as shown in the table below. After digestion, cool, transfer the contents to a 50mL volumetric flask with pure water, and dilute to the scale as shown in the table below.

[0069] Table 11. Specific solution list

[0070] Starting temperature (℃) Heating time (min) End temperature(℃) Holding time (minutes) Room temperature 5 130 8 130 5 160 8 160 5 185 25

[0071] 1.1.5 Sample solution

[0072] Take 10 bottles of medicine, open them separately and dissolve the contents, then pour them into the same plastic container and dilute to 50ml as the sample mother solution. Take 1.0mL of the sample mother solution in the digestion tank and prepare it in the same way as the "specific solution" starting from "add 7mL of nitric acid"; prepare 2 portions in parallel using the same method.

[0073] 1.1.6 Accuracy Solution

[0074] Prepare low-concentration level solution: take 1.0 mL of sample mother solution into a digestion tank, add 0.2 mL of spike solution, and prepare in the same way as the "specific solution" starting from "add 7 mL of nitric acid", with the spike concentration of 8 μg / L; prepare 3 copies in parallel using the same method.

[0075] Prepare medium concentration level solution: take 1.0mL of sample mother solution into the digestion tank, add 0.5mL of spike solution, and prepare it in the same way as the "specific solution" starting from "add 7mL of nitric acid", with the spike concentration of 20μg / L; prepare 3 copies in parallel using the same method.

[0076] Prepare high-concentration level solution: take 1.0 mL of sample mother solution into a digestion tank, add 0.75 mL of spike solution, and prepare in the same way as "specific solution" starting from "add 7 mL of nitric acid", with the spike concentration of 30 g / L; prepare 3 copies in parallel using the same method.

[0077] 1.2 Instrument parameters

[0078] Table 12. Analysis of test solution and internal standard solution injected through three-way valve in a ratio of 1:1

[0079]

[0080]

[0081]

[0082] 1.3 Verification Parameters

[0083] 1.3.1 Linear

[0084] Take the standard curve solution for sample analysis, and the instrument will automatically give a linear equation. The correlation coefficient r of the linear equation of all elements should be no less than 0.995.

[0085] 1.3.2 Limit of quantitation (LOQ)

[0086] Take a specific solution, inject 11 injections continuously for analysis, and calculate the SD of the 11 injection test results; prepare a standard solution with a value of about 10SD (adjust appropriately according to actual conditions) as the instrument LOQ solution, and calculate the recovery rate according to the linear equation. The recovery rate should be between 70% and 150%.

[0087] 1.3.3 Exclusivity

[0088] Take the specific solution for sampling and analysis, and the measured concentration of all elements should not exceed the concentration of the instrument LOQ solution.

[0089] 1.3.4 Accuracy

[0090] Take the accuracy solution for sampling and analysis, and calculate the recovery rate based on the linear equation. The recovery rate should be between 70% and 150%.

[0091] 1.4 Verification Results

[0092] 1.4.1 Linear

[0093] The correlation coefficients of the linear regression curves of all elements were greater than 0.995.

[0094] Table 13. Linear regression curves of elements

[0095] element Correlation coefficient Linear equations Al 0.9980 Y=0.000234X B 0.9999 Y=0.000015X Ba 0.9999 Y=0.024957X Si 0.9995 Y=0.000977X Sr 0.9999 Y=0.005221X Ti 0.9999 Y=0.000121X

[0096] 1.4.2 LOQ

[0097] Linear 4 solution and diluent were taken to prepare instrument LOQ solutions of 0.2μg / L, 0.5μg / L, 5.0μg / L and 50.0μg / L for sampling and analysis. The recoveries of Ti were between 70% and 150% at the concentration of 0.2μg / L, the recoveries of B, Ba and Sr were between 70% and 150% at the concentration of 0.5μg / L, the recoveries of Al were between 70% and 150% at the concentration of 5.0μg / L, and the recoveries of Si were between 70% and 150% at the concentration of 50.0μug / L.

[0098] Table 14. Recovery rate distribution table

[0099]

[0100] 1.4.3 Exclusivity

[0101] Table 15. The measured concentrations of the specific solutions are all less than the instrument L0Q distribution table

[0102]

[0103]

[0104] 1.4.4 Accuracy

[0105] Table 16. The recovery rates of the accuracy solutions are distributed between 70% and 150%.

[0106]

[0107] Table 17. Al accuracy details

[0108]

[0109] Table 18.B Accuracy Details

[0110]

[0111] Table 19. Ba accuracy details

[0112]

[0113] Table 20. Si accuracy details

[0114]

[0115]

[0116] Table 21. Sr accuracy details

[0117]

[0118] Table 22. Ti accuracy details

[0119]

[0120]

[0121] 1.5 Calculation formula

[0122] Sample LOQ (μg / g) = instrument LOQ (μg / L) × dilution factor × loading volume (Lg);

[0123] Sample result (μg / g) = instrument measured (μg / L) × dilution factor × loading volume (I / g);

[0124]

[0125] Added (μg) = spike concentration (μg / L) × added volume (L); Measured (μg) = accuracy result (μg / L) × solution volume (L);

[0126]

[0127] Table 23. Verification results details

[0128]

[0129] All validation parameters met the acceptance criteria and this method has good accuracy and can be used as a guidance document for the determination of the six elements in XQ powder injection in laboratories.

Claims

1. A method for determining the contents of six elements in a powder preparation of silver amine mesylate lactone B for injection, characterized in that The steps include: Step 1. Prepare diluent: Take 100 mL of nitric acid and dilute it to 1000 mL with water; Step 2. Prepare the spike solution: Take the standard solutions of B, Si, and Sr elements and prepare them into a solution with a concentration of 100 μg / mL according to the table below; Step 3. Prepare element stock solution: Take 100 μg / mL Al, B, Ba, Si, Sr, Ti element solution and diluent to prepare element stock solution; Step 4. Prepare standard curve solution: Take B, Ba, Sr and Ti spike solutions and diluent to prepare standard curve solution; Step 5. Take AI spike solution and diluent to prepare standard curve solution; Step 6. Take Si spike solution and diluent to prepare standard curve solution; Step 7. Prepare a specific solution: take 1.0 mL of pure water into a digestion tank, add 7 mL of nitric acid, pre-digest at 120°C for 20 min, seal after cooling, perform microwave digestion according to the table below, cool after digestion, transfer the contents to a 50 mL volumetric flask with pure water, and dilute to the mark; Step 8. Prepare sample solution: Take 10 bottles of medicine, open them respectively and dissolve the contents, then pour them into the same plastic container and dilute to 50 ml as sample mother solution; take 1.0 ml of sample mother solution into a digestion tank, and prepare it in the same way as "specific solution" starting from "add 7 ml of nitric acid"; prepare 2 portions in parallel using the same method; Step 9. Prepare the accuracy solution: Low concentration level: Take 1.0 mL of the sample mother solution in a digestion tank, add 0.2 mL of the spike solution, and prepare it in the same way as the "specific solution" starting from "add 7 mL of nitric acid", with the spike concentration of 8 μg / L; prepare 3 copies in parallel using the same method; Step 10. Prepare medium-concentration solution: Take 1.0 mL of sample stock solution and place it in a digestion tank. Add 0.5 mL of spike solution. Prepare the solution in the same way as the "specific solution" starting from "add 7 mL of nitric acid". The spike concentration is 20 μg / L. Prepare three times in parallel using the same method. Step 11. Prepare high-concentration level solution: Take 1.0 mL of sample stock solution and place it in a digestion tank. Add 0.75 mL of spike solution. Prepare the solution in the same way as the "specific solution" starting from "add 7 mL of nitric acid". The spike concentration is 30 g / L. Prepare three times in parallel using the same method.

2. The method for determining the contents of six elements in the silver amine lactone mesylate B powder injection preparation for injection according to claim 1, characterized in that: The following steps are also included: Step 12. Verify linear parameters: Take the standard curve solution for analysis, and the instrument will automatically give a linear equation. The correlation coefficient r of the linear equation of all elements should be no less than 0.

995. Step 13. Verify the limit of quantitation LOQ: Take a specific solution, inject 11 injections continuously for analysis, and calculate the SD of the test results of the 11 injections; prepare a standard solution with a limit of 10SD as the instrument LOQ solution, and calculate the recovery rate according to the linear equation. The recovery rate should be between 70% and 150%; Step 14. Verify specificity: Take a sample of the specific solution for analysis. The concentration of all elements measured should not exceed the concentration of the instrument LOQ solution. Step 15. Verify accuracy: Take the accuracy solution for analysis and calculate the recovery rate according to the linear equation. The recovery rate should be between 70% and 150%.

3. The method for determining the contents of six elements in the silver amine lactone mesylate B powder injection preparation for injection according to claim 2, characterized in that: The calculation formula for the verification accuracy in step 15 is: Sample LOQ (μg / g) = instrument LOQ (μg / L) × dilution factor × loading volume (Lg); Sample result (μg / g) = instrument measured (μg / L) × dilution factor × loading volume (I / g); Added (μg) = spike concentration (μg / L) × added volume (L); Measured (μg) = accuracy result (μg / L) × solution volume (L);

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