Ultraviolet spectrophotometer determination method for content of silicon element in protease medicine

Abstract

The invention relates to an ultraviolet spectrophotometer determination method for the content of silicon elements in protease drugs, which comprises the following steps: S1, sample pretreatment: pretreatment of a test sample and pretreatment of a labeled test sample; s2, making a standard curve; s3, sample determination; and S4, calculating a result. The pretreatment steps of the test sample and the labeled test sample are detailed and consistent, variation in the sample preparation process is reduced, the comparability between samples is ensured, and the method has the advantages of high stability, high repeatability, high accuracy, simplicity and convenience in operation and wide applicability.

Description

Technical Field

[0001] The present invention belongs to the technical field of protease drugs, and particularly relates to a method for determining the silicon element content in protease drugs by ultraviolet spectrophotometry. Background Art

[0002] In protease drugs, the determination of silicon element is of great significance for ensuring the quality and safety of drugs. Silicon element may come from catalysts, raw materials, water or excipients used in the drug production process, as well as impurities generated by the interaction with production equipment or packaging systems. Therefore, the accurate determination of silicon element helps to control the elemental impurities in drugs to avoid possible toxicity risks.

[0003] At present, the detection equipment for silicon detection is expensive and has high maintenance costs. At the same time, there is also a serious interference in sample analysis, which affects the accuracy of test results. Summary of the Invention

[0004] (I) Object of the Invention

[0005] In order to overcome the above deficiencies, the object of the present invention is to provide a method for determining the silicon element content in protease drugs by ultraviolet spectrophotometry to solve the above technical problems.

[0006] (II) Technical Solution

[0007] To achieve the above object, the technical solution provided by the present application is as follows:

[0008] A method for determining the silicon element content in protease drugs by ultraviolet spectrophotometry, comprising the following steps:

[0009] S1 Sample pretreatment, including the pretreatment of the test sample and the pretreatment of the spiked test sample;

[0010] S2 Standard curve preparation: Add 0 ml, 0.25 ml, 0.5 ml, 1 ml, 2 ml of silicon standard solution to a 25 ml volumetric flask, and successively add sulfuric acid and ammonium molybdate, slightly heat, then add oxalic acid solution and ammonium ferrous sulfate solution, dilute with water, and detect at a wavelength of 680 nm;

[0011] S3 Sample determination: Take 2.5 ml of the pretreatment solution of the test sample and the spiked test sample and add them to a 25 ml volumetric flask, successively add 2.5 ml of 0.5 mol / sulfuric acid; 2.5 ml of ammonium molybdate (50 g / L), place in a water bath at 35 °C for 5 min, then add 2.5 mL of oxalic acid solution and shake well, then add 2.5 mL of ammonium ferrous sulfate solution (60 g / L), dilute with water to the scale and let stand for 10 min, and detect at a wavelength of 680 nm;

[0012] S4 Result calculation.

[0013] Preferably, the pretreatment of the test sample includes dissolving the test sample and making up the volume to the scale, taking 2.5 ml of the dissolved test sample, adding 2.5 ml of trichloroacetic acid solution, mixing well, standing for more than 30 minutes, and then centrifuging.

[0014] Preferably, the test sample is 0.04 g, the volume of the volumetric flask is 5 ml, the concentration of the trichloroacetic acid solution is 10 - 15%, the centrifugation speed > 4000 r / min, and the centrifugation time is 5 - 10 min.

[0015] Preferably, the pretreatment of the spiked test sample includes dissolving the test sample by adding water, adding 0.4 ml of 100 μg / ml silicon element standard stock solution, dissolving by adding water and making up the volume to the scale, taking 2.5 ml of the above solution, adding 2.5 ml of 10 - 15% trichloroacetic acid solution, mixing well, standing for 30 minutes, and then centrifuging, the centrifugation speed > 4000 r / min, and the centrifugation time is 5 - 10 min.

[0016] Preferably, the concentration of the silicon standard solution in S2 is 10 μg / ml, add 2.5 ml of sulfuric acid with a concentration of 0.5 mol / L, add 2.5 ml of ammonium molybdate with a concentration of 50 g / L, after slightly heating, then add 2.5 mL of oxalic acid solution and shake well, and then add 2.5 mL of ammonium ferrous sulfate solution with a concentration of 60 g / L.

[0017] Preferably, the way of slightly heating in S2 is to place it in a 35°C water bath for 5 min.

[0018] Preferably, it needs to stand for 8 - 15 minutes before wavelength detection in S2.

[0019] Beneficial effects:

[0020] The pretreatment steps of the test sample and the spiked test sample of the present invention are detailed and consistent, which helps to reduce the variation in the sample preparation process, ensures the comparability between samples, and has the advantages of high stability, high repeatability, high accuracy, simple operation and wide applicability. Description of the drawings

[0021] Figure 1 It is a schematic diagram of the Si content standard curve of the present invention. Specific embodiments

[0022] To make the purpose, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that these descriptions are exemplary and are not intended to limit the scope of the present invention. In addition, in the following description, the descriptions of well-known structures and technologies are omitted to avoid unnecessarily confusing the concepts of the present invention.

[0023] A method for determining the silicon element content in protease drugs provided by the present invention is characterized by comprising the following steps:

[0024] S1 Sample pretreatment, including test sample pretreatment and spiked test sample pretreatment;

[0025] Test sample pretreatment: Weigh 0.04 g of the test sample precisely into a 5-ml volumetric flask, dissolve it and make up the volume to the mark. Take 2.5 ml of the above solution, add 2.5 ml of 10 - 15% trichloroacetic acid solution, mix well, let stand for more than 30 minutes, then centrifuge, with a centrifuge speed > 4000 r / min and a centrifugation time of 5 - 10 min.

[0026] Spiked test sample pretreatment: Weigh 0.04 g of the test sample precisely into a 5-ml volumetric flask, add appropriate amount of water to dissolve it, then add 0.4 ml of 100 μg / ml silicon element standard stock solution, add water to dissolve and make up the volume to the mark. Take 2.5 ml of the above solution, add 2.5 ml of 10 - 15% trichloroacetic acid solution, mix well, let stand for 30 minutes, then centrifuge, with a centrifuge speed > 4000 r / min and a centrifugation time of 5 - 10 min.

[0027] S2 Standard curve preparation: Respectively take 0 ml, 0.25 ml, 0.5 ml, 1 ml, 2 ml of 10 μg / ml silicon standard solution into 25-ml volumetric flasks, and successively add 2.5 ml of 0.5 mol / L sulfuric acid; 2.5 ml of 50 g / L ammonium molybdate, place in a water bath at 35°C for 5 min, then add 2.5 mL of oxalic acid solution and shake well, then add 2.5 mL of 60 g / L ammonium ferrous sulfate solution, dilute with water to the mark and let stand for 10 min, and detect at a wavelength of 680 nm.

[0028] S3 Sample determination: Take 2.5 ml of the test sample and spiked test sample pretreatment solutions into 25-ml volumetric flasks, and successively add 2.5 ml of 0.5 mol / sulfuric acid; 2.5 ml of 50 g / L ammonium molybdate, place in a water bath at 35°C for 5 min, then add 2.5 mL of oxalic acid solution and shake well, then add 2.5 mL of 60 g / L ammonium ferrous sulfate solution, dilute with water to the mark and let stand for 10 min, and detect at a wavelength of 680 nm.

[0029] S4 Result calculation.

[0030] 1. The linear equation is y = 0.0125x - 0.0002, R 2 = 0.9999, the content on the Y-axis (μg), the X-axis is the absorbance value, and the R value is the linear regression coefficient. For details, see Figure 1 , indicating that the method has good linearity, meets the requirements of quantitative detection, has high accuracy, consistent sample preparation, and good instrument stability.

[0031] 2. Six spiked test samples were set up, and the Si content was calculated using a linear equation, labeled as a, b, c, d, e, and f respectively. The following are the repeatability test data.

[0032]

[0033] The data of the spiked repeatability test showed that the RSD value was 0.42%, < 0.5%, indicating that the method had good stability and high reliability. RSD is an index to measure the degree of data dispersion. It is the ratio of the standard deviation (SD) to the average of the measured values, usually expressed as a percentage. The RSD value of 0.42% is much lower than 0.5%, which indicates high stability and high reliability. Generally speaking, an RSD value less than 0.5% is a very good result, indicating that the analytical method used performs excellently in terms of repeatability and can be used for accurate and reliable quantitative analysis.

[0034] 3. Recovery rate of the spiked test

[0035]

[0036]

[0037] The recovery rate of the spiked test was 99.6%, within the range of 95% - 105%, indicating that the method had high accuracy; the RSD of the recovery rate was 0.42%, indicating good repeatability of the spiked recovery rate.

[0038] Therefore, the determination method of the present invention showed excellent linear relationship, high accuracy, high repeatability, and high stability, and was suitable for quantitative analysis of silicon content.

[0039] It should be noted that in this article, relational terms such as first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variant thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements not only includes those elements, but also includes other elements not expressly listed, or also includes elements inherent in such process, method, article or device. Without further limitation, an element defined by the statement "including a..." does not exclude the existence of additional identical elements in the process, method, article or device including the said element.

[0040] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the various embodiments of the present invention.

Claims

1. A method for determining the silicon content in protease drugs by ultraviolet spectrophotometry, characterized in that: The following steps are involved: S1 sample pretreatment, including sample pretreatment and spiked sample pretreatment; To prepare the S2 standard curve, add 0ml, 0.25ml, 0.5ml, 1ml and 2ml of silicon standard solution to a 25ml volumetric flask, and add sulfuric acid and ammonium molybdate in sequence, heat slightly, then add oxalic acid solution and ammonium ferrous sulfate solution, dilute with water and detect at a wavelength of 680nm; For S3 sample determination, take 2.5 ml of the sample and the pretreatment solution of the spiked sample into a 25 ml volumetric flask, add 2.5 ml of 0.5 mol / sulfuric acid and 2.5 ml of ammonium molybdate (50 g / L) in turn, place in a 35°C water bath for 5 min, then add 2.5 ml of oxalic acid solution and shake thoroughly, then add 2.5 ml of ammonium ferrous sulfate solution (60 g / L), dilute to the scale with water and place for 10 min, and detect at a wavelength of 680 nm; S4 result calculation.

2. The method for determining the silicon content in a protease drug by ultraviolet spectrophotometry according to claim 1, characterized in that: The sample pretreatment includes dissolving the sample and fixing the volume to the mark, taking 2.5 ml of the dissolved sample, adding 2.5 ml of trichloroacetic acid solution, mixing, standing for more than 30 minutes, and then centrifuging.

3. The method for determining the silicon content in a protease drug by ultraviolet spectrophotometry according to claim 2, characterized in that: The test sample is 0.04 g, the capacity of the volumetric flask is 5 ml, the concentration of the trichloroacetic acid solution is 10-15%, the centrifugal speed is >4000 r / min, and the centrifugal time is 5-10 min.

4. The method for determining the silicon content in a protease drug by ultraviolet spectrophotometry according to claim 1, characterized in that: The pretreatment of the spiked test sample includes dissolving the test sample in water, adding 0.4 ml of 100 μg / ml silicon element standard stock solution, dissolving in water and diluting to the scale, taking 2.5 ml of the above solution, adding 2.5 ml of 10-15% trichloroacetic acid solution, mixing and standing for 30 minutes and then centrifuging, the centrifugal speed is greater than 4000 r / min, and the centrifugal time is 5-10 min.

5. The method for determining the silicon content in a protease drug by ultraviolet spectrophotometry according to claim 1, characterized in that: The concentration of the silicon standard solution in S2 is 10 μg / ml. 2.5 ml of 0.5 mol / L sulfuric acid and 2.5 ml of 50 g / L ammonium molybdate are added. After slight heating, 2.5 ml of oxalic acid solution is added, shaken thoroughly, and then 2.5 ml of 60 g / L ammonium ferrous sulfate solution is added.

6. The method for determining the silicon content in a protease drug by ultraviolet spectrophotometry according to claim 1, characterized in that: The method of slightly heating S2 is to place it in a 35° C. water bath for 5 minutes.

7. The method for determining the silicon content in a protease drug by ultraviolet spectrophotometry according to claim 1, characterized in that: The S2 needs to be left for 8-15 minutes before wavelength detection.

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