Florfenicol particle size analysis method

Abstract

The invention belongs to the technical field of biological medicines, and particularly discloses a Florfenicol particle size analysis method, which is carried out by adopting a particle size analyzer, and is characterized by comprising the following steps: uniformly shaking a Florfenicol raw material medicine sample, putting the sample into a test tube, adding a dispersing agent into the test tube, carrying out vortex treatment until the sample is uniformly dispersed, preparing a sample suspension, and carrying out freeze drying on the sample suspension to obtain the Florfenicol particle size analysis product. The sample suspension is taken and placed in a particle size analyzer to be subjected to background testing, after the background testing is completed, the sample suspension is shaken up, the shaken-up sample suspension is added into a dispersing pool of the particle size analyzer, the particle size analyzer is used for carrying out wet-method particle size determination, and an analysis result is obtained. The invention provides a Florfenicol particle size analysis method, which is used for providing a stable and accurate measurement method for the measurement of the particle size of Florfenicol.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and in particular relates to a Florfenicol particle size analysis method. Background Art

[0002] Florfenicol is a synthetic monofluoro derivative of thiamphenicol and an antibiotic drug (veterinary drug). It can diffuse into bacterial cells through fat solubility and mainly acts on the 50s subunit of the bacterial 70s ribosome, inhibiting transpeptidase, hindering the growth of peptidase, inhibiting the formation of peptide chains, thereby preventing protein synthesis and achieving antibacterial purposes.

[0003] Florfenicol has a broad antibacterial spectrum, including various Gram-positive and Gram-negative bacteria and mycoplasmas, etc. Sensitive bacteria include Haemophilus influenzae of cattle and pigs, Shigella dysenteriae, Salmonella, Escherichia coli, Pneumococcus, Haemophilus influenzae, Streptococcus, Staphylococcus aureus, Chlamydia, Leptospira, Rickettsia, etc.

[0004] Florfenicol is rapidly absorbed orally, widely distributed, has a long half-life, high blood concentration, and long blood drug maintenance time. The Florfenicol API produced by our company needs to be used in the production of Florfenicol preparations, so the particle size of Florfenicol API needs to be strictly controlled. Currently, there is no method for determining and analyzing the particle size of Florfenicol. Summary of the invention

[0005] The purpose of the present invention is to overcome the above problems existing in the prior art and provide a Florfenicol particle size analysis method for providing a stable and accurate determination method for the Florfenicol particle size.

[0006] In order to achieve the above technical objectives and the above technical effects, the present invention is implemented through the following technical solutions:

[0007] A method for analyzing the particle size of Florfenicol is carried out using a particle size analyzer, wherein the method comprises the following steps:

[0008] After the Florfenicol API sample is shaken, it is placed in a test tube, a dispersant is added to the test tube, and a sample suspension is prepared after vortexing until the sample is evenly dispersed. The sample suspension is taken to a particle size analyzer for background test. After the background test is completed, the sample suspension is shaken, and the shaken sample suspension is added to the dispersion tank of the particle size analyzer. The particle size analyzer is used to perform wet particle size measurement to obtain analysis results;

[0009] During the wet particle size determination process, the relevant measurement parameters of the particle size analyzer are as follows:

[0010] Sample suspension standing time: 0-30min;

[0011] Dispersion time: 5-10min;

[0012] Stirring speed: 1600-2000rpm;

[0013] Shading degree: 10-20%.

[0014] Furthermore, among the relevant measurement parameters of the particle size analyzer, the standing time of the sample suspension is 15 minutes.

[0015] Furthermore, among the relevant measurement parameters of the particle size analyzer, the dispersion time is 10 minutes.

[0016] Furthermore, among the relevant measurement parameters of the particle size analyzer, the stirring speed of the disperser in the particle size analyzer is 1800 rpm.

[0017] Furthermore, among the relevant measurement parameters of the particle size analyzer, the shading degree is 10%.

[0018] Furthermore, in the wet particle size determination process, the relevant measurement parameters of the particle size analyzer also include:

[0019] Delay time before measurement: 0-60S;

[0020] Delay before focusing: 300S;

[0021] Number of measurements: 3 times.

[0022] Furthermore, the dispersant is Miglyol 812N.

[0023] Furthermore, in the sample suspension, the mass volume ratio of the sample to the dispersant is 6-15 mg / mL.

[0024] Furthermore, the refractive index of the dispersant is 1.2-1.5.

[0025] Furthermore, the refractive index of the dispersant is 1.45.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] 1. The present invention provides a Florfenicol particle size analysis method, which is used to provide a stable and accurate determination method for the Florfenicol particle size.

[0028] 2. In the present invention, Miglyol 812N is used as a dispersant, and the sample is dispersed evenly without agglomeration, indicating that Miglyol 812N as a dispersant has a certain specificity for the Florfenicol particle size analysis.

[0029] 3. In the present invention, by determining the parameter ranges of the sample suspension standing time, dispersion time, stirring speed and shading degree, the particle size is measured within this measurement parameter range, and the measurement results are repeatable, indicating that the Florfenicol particle size analysis method provided by the present invention is suitable for the determination of Florfenicol particle size.

[0030] 4. The Florfenicol particle size analysis method provided in the present invention has good specificity, precision and durability, and is suitable for the detection of Florfenicol particle size. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] The drawings described herein are used to provide a further understanding of the present invention and constitute a part of this application. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the drawings:

[0032] Figure 1 It is a sample particle size distribution curve diagram under different sample suspension standing times provided by the present invention; Figure 1 In the figure, the x-axis is the particle size; curve 11 is: differential distribution (%), which refers to the percentage of each particle size; curve 12 is: cumulative distribution (%), which refers to the cumulative percentage of particle size.

[0033] Figure 2 It is a sample particle size distribution curve diagram under different dispersion times provided by the present invention; Figure 2 In the figure, the x-axis is the particle size; curve 21 is: differential distribution (%), which refers to the percentage of each particle size; curve 22 is: cumulative distribution (%), which refers to the cumulative percentage of particle size.

[0034] Figure 3 It is a sample particle size distribution curve diagram under different shading degrees provided by the present invention; Figure 3 In the figure, the x-axis is the particle size; curve 31 is: differential distribution (%), which refers to the percentage of each particle size; curve 32 is: cumulative distribution (%), which refers to the cumulative percentage of particle size.

[0035] Figure 4 It is a sample particle size distribution curve diagram under different stirring speeds provided by the present invention; Figure 4 In the figure, the x-axis is the particle size; curve 41 is: differential distribution (%), which refers to the percentage of each particle size; curve 42 is: cumulative distribution (%), which refers to the cumulative percentage of particle size.

[0036] Figure 5 It is a sample particle size distribution curve diagram provided in Comparative Example 1 of the present invention when the disperser speed is 2200 rpm; Figure 5 In the figure, the x-axis is the particle size; curve 51 is: differential distribution (%), which refers to the percentage of each particle size; curve 52 is: cumulative distribution (%), which refers to the cumulative percentage of particle size.

[0037] Figure 6 It is a sample particle size distribution curve diagram provided in Comparative Example 2 of the present invention when the dispersion time is 15 min; Figure 6 In the figure, the x-axis is the particle size; curve 61 is: differential distribution (%), which refers to the percentage of each particle size; curve 62 is: cumulative distribution (%), which refers to the cumulative percentage of particle size.

[0038] Figure 7 is a particle size distribution curve diagram of 6 samples tested by analyst A provided by the present invention; Figure 7 In the figure, the x-axis is the particle size; curve 71 is: differential distribution (%), which refers to the percentage of each particle size; curve 72 is: cumulative distribution (%), which refers to the cumulative percentage of particle size.

[0039] Figure 8 is a sample particle size distribution curve diagram of 6 samples tested by analyst B provided by the present invention; Figure 8 In the figure, the x-axis is the particle size; curve 81 is: differential distribution (%), which refers to the percentage of each particle size; curve 82 is: cumulative distribution (%), which refers to the cumulative percentage of particle size.

[0040] Fig. 9 It is a sample particle size distribution curve diagram of 12 samples of analysts A and B provided by the present invention; Fig. 9 In the figure, the x-axis is the particle size; curve 91 is: differential distribution (%), which refers to the percentage of each particle size; curve 92 is: cumulative distribution (%), which refers to the cumulative percentage of particle size. DETAILED DESCRIPTION

[0041] The following will be combined with the accompanying drawings to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. 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.

[0042] The present invention provides a Florfenicol particle size analysis method, which is performed using a particle size analyzer. The analysis method comprises the following steps:

[0043] S1: After the sample is shaken, it is placed in a test tube, a dispersant is added to the test tube, and a sample suspension is prepared after vortexing until the sample is evenly dispersed:;

[0044] In step S1, the dispersant is Miglyol 812N; the refractive index of the dispersant is 1.45;

[0045] In the sample suspension, the mass volume ratio of sample to dispersant is 6-15 mg / mL;

[0046] S2: Take the sample suspension for background test. After the background test is completed, shake the sample suspension, add the shaken sample suspension into the dispersion tank, perform wet particle size determination, and obtain analysis results.

[0047] In step S2, during the wet particle size determination process, the determination parameters are as follows:

[0048] Sample suspension standing time: 0-30min;

[0049] Dispersion time: 5-10min;

[0050] Stirring speed: 1600-2000rpm;

[0051] Shading degree: 10-20%;

[0052] Specifically, the measurement parameters also include:

[0053] Delay time before measurement: 0-60S;

[0054] Delay before focusing: 300S;

[0055] Number of measurements: 3 times.

[0056] The Florfenicol particle size analysis method provided by the present invention is further explained and demonstrated in combination with specific examples and experimental results. In the following examples, unless otherwise specified, all raw materials, instruments and equipment can be obtained commercially.

[0057] Example 1

[0058] This embodiment provides a method for analyzing the particle size of Florfenicol, which is performed using a particle size analyzer, and the relevant measurement parameters are as follows:

[0059]

[0060]

[0061] The relevant setting parameters are as follows:

[0062] type parameter Sample suspension standing time 15min Spread out time 10min Stirring speed 1800rpm Shading 10％

[0063] The specific operation process is as follows: gently shake the Florfenicol API sample, place 100 mg of the sample in a 25 mL stoppered test tube, add 10 mL of dispersant Miglyol 812N, vortex until the Florfenicol API sample is evenly dispersed, and prepare a sample suspension; first perform a background test, after the background test is completed, shake the sample suspension, add it to the dispersion pool of the particle size analyzer, make the shading degree 10%, start measurement, and report the particle size measurement data.

[0064] Example 2

[0065] This embodiment provides a Florfenicol particle size analysis method, which is performed using a particle size analyzer. Compared with Example 1, in this embodiment, the shading degree is 15%, and the remaining parameters and operation procedures are the same as those in Example 1.

[0066] Example 3

[0067] This comparative example provides a method for analyzing the particle size of Florfenicol using a particle size analyzer. Compared with Example 1, in this example, the shading degree is 20%, and the other parameters and operation procedures are the same as those in Example 1.

[0068] Example 4

[0069] This embodiment provides a Florfenicol particle size analysis method, which is performed using a particle size analyzer. Compared with Example 1, in this embodiment, the sample suspension is left to stand for 0 min, and the remaining parameters and operation procedures are the same as those in Example 1.

[0070] Example 5

[0071] This embodiment provides a Florfenicol particle size analysis method, which is performed using a particle size analyzer. Compared with Example 1, in this embodiment, the sample suspension is allowed to stand for 30 minutes, that is, the sample is measured without being allowed to stand. The remaining parameters and operating procedures are the same as in Example 1.

[0072] Example 6

[0073] This embodiment provides a Florfenicol particle size analysis method, which is performed using a particle size analyzer. Compared with Example 1, in this embodiment, the dispersion time is 0 min, and the other parameters and operation procedures are the same as those in Example 1.

[0074] Example 7

[0075] This embodiment provides a Florfenicol particle size analysis method, which is performed using a particle size analyzer. Compared with Example 1, in this embodiment, the dispersion time is 5 minutes, and the other parameters and operation procedures are the same as those in Example 1.

[0076] Example 8

[0077] This embodiment provides a Florfenicol particle size analysis method, which is performed using a particle size analyzer. Compared with Example 1, in this embodiment, the stirring speed is 1600 rpm, and the other parameters and operation procedures are the same as those in Example 1.

[0078] Example 9

[0079] This embodiment provides a Florfenicol particle size analysis method, which is performed using a particle size analyzer. Compared with Example 1, in this embodiment, the stirring speed is 2000 rpm, and the other parameters and operation procedures are the same as those in Example 1.

[0080] Comparative Example 1

[0081] This comparative example provides a Florfenicol particle size analysis method, which is performed using a particle size analyzer. Compared with Example 1, in this comparative example, the stirring speed is 2200 rpm, and the other parameters and operation procedures are the same as those in Example 1.

[0082] Comparative Example 2

[0083] This comparative example provides a Florfenicol particle size analysis method, which is performed using a particle size analyzer. Compared with Example 1, in this comparative example, the dispersion time is 15 minutes, and the other parameters and operation procedures are the same as those in Example 1.

[0084] Experimental Example 1

[0085] Data analysis was performed on the particle size test results of the Florfenicol bulk drug samples in Examples 1-9 and Comparative Examples 1-2. During the data analysis, the percentage of particles with a particle size of <10 μm, <25 μm, <100 μm, <200 μm, and <300 μm in the samples, and the RSD (%) of the particle size values ​​of D(10), D(50), and D(90) should all be no more than 20%, that is, the acceptance standard should be no more than 20%.

[0086] The specific analysis results are shown in Table 1, Table 2, Table 3, Table 4, Table 5 and Table 6.

[0087] Table 1 Particle size test results of different sample suspension standing time

[0088]

[0089]

[0090] Table 1 shows the particle size test results obtained in Example 1, Example 3 and Example 4 at different sample suspension standing times. The specific distribution chart is as follows: Figure 1 As shown, the results show that under different sample suspension standing times, the particle size test results are all within an acceptable range, and according to the data in the table, in Example 1, that is, when the sample suspension is standing for 15 minutes, the proportion of <200 μm and <300 μm are both relatively large, indicating good durability under the test conditions.

[0091] Table 2 Particle size test results at different dispersion times

[0092]

[0093] Table 2 shows the particle size test results of Example 5, Example 6 and Example 1 at different dispersion times. The specific distribution chart is as follows: Figure 2 As shown, the results show that under different dispersion times, the particle size test results are all within an acceptable range, indicating that when the dispersion time is 5-10min, it is a suitable test condition, and according to the test results, in Example 1, the proportion of <300μm is relatively large, indicating that under this test condition, it has good durability;

[0094] Table 3 Particle size test results under different shading degrees

[0095]

[0096]

[0097] Table 3 shows the particle size test results of Example 2, Example 1, Example 2 and Example 3 at different shading degrees. The specific distribution chart is as follows: Figure 3 As shown, the results show that under different shading degrees, the particle size test results are within the acceptable range, indicating that when the shading degree is 5-20%, it is a suitable test condition, and according to the test results, in Example 1, the proportion of <300μm is relatively large, indicating that under this test condition, it has good durability;

[0098] Table 4 Particle size test results at different disperser stirring speeds

[0099]

[0100] Table 4 shows the particle size test results of Example 2, Example 1, Example 2 and Example 3 at different shading degrees. The specific distribution chart is as follows: Figure 4As shown, the results show that under different shading degrees, the particle size test results are within the acceptable range, indicating that when the stirring speed of the disperser is 1600-2000rpm, it is a suitable test condition, and according to the test results, in Example 1, the proportion of <300μm is relatively large, indicating that under this test condition, it has good durability;

[0101] Table 5 Particle size test results of comparative example 1 at a disperser speed of 2200 rpm

[0102]

[0103] Table 5 shows the particle size test results provided in Comparative Example 1, with the disperser speed set at 2200 rpm. The specific distribution chart is as follows: Figure 5 As shown, compared with the data in Table 4, it can be seen that due to the high rotation speed, bubbles will be generated in the disperser, which will affect the test results;

[0104] Table 6 Particle size test results when the dispersion time is 15min provided in Comparative Example 2

[0105]

[0106] Table 6 shows the particle size test results when the dispersion time is 15 min provided in Example 2. The specific distribution chart is as follows: Figure 6 As shown, compared with the data in Table 2, it can be seen that if the dispersion time is too long, the particle size test result will be too large due to particle agglomeration;

[0107] Based on the above analysis, under the following parameters, namely: sample suspension standing time: 0-30 min; dispersion time: 5-10 min; stirring speed: 1600-2000 rpm; shading degree: 5-20%, the relative standard deviation of the Florfenicol particle size test results at this time meets the requirements of the acceptable standard. Therefore, the Florfenicol particle size analysis method provided by the present invention has good specificity, precision and durability.

[0108] Experimental Example 2

[0109] This experimental example is to conduct an experiment with reference to the experimental parameters provided in Example 1, to verify that the Florfenicol particle size analysis method provided by the present invention has good repeatability.

[0110] In this experimental example, two analysts conducted the experiment according to the experimental parameters provided in Example 1. The two analysts were marked as A and B respectively. Each analyst prepared 6 groups of samples. The obtained Florfenicol particle size test results are shown in Tables 7, 8 and 9.

[0111] Table 7 Florfenicol particle size test results of analyst A

[0112]

[0113]

[0114] According to the analysis results in Table 7 and Figure 7 It can be seen from the distribution diagram that among the 6 samples tested by analyst A, the percentage of particles <10μm, <25μm, <100μm, <200μm, <300μm and the particle size values ​​of D(10), D(50), and D(90) are all less than 20%, that is, the repeatability of the particle size test results is within an acceptable range;

[0115] Table 8 Florfenicol particle size test results of analyst B

[0116]

[0117] According to the analysis results in Table 8 and Figure 8 It can be seen from the distribution diagram that among the 6 samples tested by analyst B, the percentage of particles with a particle size of less than 10 μm, less than 25 μm, less than 100 μm, less than 200 μm, and the RSD (%) of the particle size values ​​of D(10), D(50), and D(90) are all less than 20%, that is, the repeatability of the particle size test results is within an acceptable range;

[0118] Table 9 Florfenicol particle size test results of analysts A and B

[0119]

[0120]

[0121] According to the analysis results in Table 9 and Fig. 9 It can be seen from the distribution diagram that for a total of 12 sample particle size tests conducted by analysts A and B, the percentage of particles <10 μm, <25 μm, <100 μm, <200 μm, <300 μm and the RSD (%) of the particle size values ​​of D(10), D(50), and D(90) were all less than 20%, and the intermediate precision results were within an acceptable range.

[0122] Therefore, this analytical method has good intermediate precision.

[0123] This experimental example proves that the Florfenicol particle size analysis method provided by the present invention has good repeatability and is suitable for the detection of the particle size of Florfenicol.

[0124] Experimental Example 3

[0125] This experimental example verifies the specificity of the Florfenicol particle size analysis method provided by the present invention in the particle size determination of Florfenicol raw materials, specifically:

[0126] S31: Preparation of sample suspension

[0127] Gently shake the Florfenicol API sample, place 100 mg of the sample in a 25 mL stoppered test tube, add 10 mL of dispersant Miglyol 812N, and vortex until the Florfenicol API sample is evenly dispersed to prepare a sample suspension;

[0128] S32: Dispersion detection

[0129] Visually inspect the sample suspension to preliminarily determine the sample suspension effect. If the sample is visually dispersed evenly without agglomeration, it can be preliminarily judged as qualified.

[0130] S33: Incompatibility with dispersants

[0131] Under the condition of shading degree of about 15%, the RSD (%) of shading degree at 5 different dispersion times (0min, 5min, 10min, 20min, 30min) was measured, among which, the RSD (%) should be no more than 20% to meet the standard.

[0132] The test results are shown in Table 10.

[0133] Table 10 Validation of the specificity of the Florfenicol particle size analysis method for the determination of the particle size of the Florfenicol API

[0134]

[0135] Among them, according to preliminary judgment, the samples at 5 different dispersion times are evenly dispersed in the dispersant without agglomeration; from the verification results in Table 10, the RSD (%) of the shading degree tested at 5 different times is no more than 20%, and all the verification results are within an acceptable range; therefore, this method has good specificity.

[0136] The invention provides a Florfenicol particle size analysis method, which has good specificity, precision and durability and can stably and accurately measure the particle size of the Florfenicol raw material medicine.

[0137] In the description of this specification, the description with reference to the terms "one embodiment", "example", "specific example", etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.

[0138] The above shows and describes the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments, and the above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, and these changes and improvements all fall within the scope of the present invention to be protected.

Claims

1. A method for analyzing the particle size of Florfenicol using a particle size analyzer, characterized in that: The analytical method comprises the following steps: After the Florfenicol API sample is shaken, it is placed in a test tube, a dispersant is added to the test tube, and a sample suspension is prepared after vortexing until the sample is evenly dispersed. The sample suspension is taken to a particle size analyzer for background test. After the background test is completed, the sample suspension is shaken, and the shaken sample suspension is added to the dispersion tank of the particle size analyzer. The particle size analyzer is used to perform wet particle size measurement to obtain analysis results; During the wet particle size determination process, the relevant measurement parameters of the particle size analyzer are as follows: Sample suspension standing time: 0-30min; Dispersion time: 5-10min; Stirring speed: 1600-2000rpm; Shading degree: 5-20%.

2. A Florfenicol particle size analysis method according to claim 1, characterized in that: Among the relevant measurement parameters of the particle size analyzer, the sample suspension standing time is 15 minutes.

3. A Florfenicol particle size analysis method according to claim 1, characterized in that: Among the relevant measurement parameters of the particle size analyzer, the dispersion time is 10 minutes.

4. A Florfenicol particle size analysis method according to claim 1, characterized in that: Among the relevant measurement parameters of the particle size analyzer, the stirring speed of the disperser in the particle size analyzer is 1800 rpm.

5. A Florfenicol particle size analysis method according to claim 1, characterized in that: Among the relevant measurement parameters of the particle size analyzer, the shading degree is 10%.

6. A Florfenicol particle size analysis method according to claim 1, characterized in that: In the wet particle size determination process, the relevant measurement parameters of the particle size analyzer also include: Delay time before measurement: 0-60S; Delay before focusing: 300S; Number of measurements: 3 times.

7. A Florfenicol particle size analysis method according to claim 1, characterized in that: The dispersant is Miglyol 812N.

8. A Florfenicol particle size analysis method according to claim 1, characterized in that: In the sample suspension, the mass volume ratio of the sample to the dispersant is 6-15 mg / mL.

9. A Florfenicol particle size analysis method according to claim 1, characterized in that: The refractive index of the dispersant is 1.2-1.

5.

10. A Florfenicol particle size analysis method according to claim 1, characterized in that: The refractive index of the dispersant is 1.45.

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