阅读说明：本技术 地塞米松磷酸钠生产过程中酯化阶段反应液中地塞米松的检测方法 (Method for detecting dexamethasone in reaction liquid in esterification stage in production process of dexamethasone sodium phosphate ) 是由 侯丽英 杨剑波 于 2020-12-18 设计创作，主要内容包括：本发明属于检测方法技术领域,具体涉及一种地塞米松磷酸钠生产过程中酯化阶段反应液中地塞米松的检测方法,包括如下步骤：(1)系统适用性溶液的制备；(2)供试品溶液的制备：取地塞米松磷酸酯酯化反应液0.2ml,加入浓盐酸0.1ml和水1ml,摇匀,再加入流动相稀释至10ml,混匀得到供试品溶液；(3)空白溶液为流动相10ml；(4)采用高效液相色谱法进行检测供试品溶液中塞米松磷酸酯；所述流动相的配制过程为：三乙胺溶液45ml、甲醇47.5ml和乙腈7.5ml混合均匀得到流动相。本发明方法能够有检测出地塞米松磷酸钠酯化反应中物料地塞米松是否反应完全,控制产品中杂质地塞米松的含量,提高收率。(The invention belongs to the technical field of detection methods, and particularly relates to a method for detecting dexamethasone in reaction liquid in an esterification stage in a production process of dexamethasone sodium phosphate, which comprises the following steps: (1) preparing a system applicability solution; (2) preparation of a test solution: taking 0.2ml of dexamethasone phosphate esterification reaction liquid, adding 0.1ml of concentrated hydrochloric acid and 1ml of water, shaking up, adding a mobile phase to dilute to 10ml, and mixing uniformly to obtain a test solution; (3) the blank solution is 10ml of mobile phase; (4) detecting the cefazedone phosphate in the test sample solution by adopting a high performance liquid chromatography; the preparation process of the mobile phase comprises the following steps: 45ml of triethylamine solution, 47.5ml of methanol and 7.5ml of acetonitrile are mixed uniformly to obtain a mobile phase. The method can detect whether the dexamethasone reacts completely in the esterification reaction of the dexamethasone sodium phosphate, control the content of dexamethasone serving as an impurity in the product and improve the yield.)

1. The method for detecting dexamethasone in reaction liquid in the esterification stage in the production process of dexamethasone sodium phosphate is characterized by comprising the following steps:

(1) preparation of system suitability solution:

firstly, preparing a dexamethasone phosphate stock solution by using a dexamethasone phosphate reference substance and a methanol solvent;

then, preparing dexamethasone stock solution by using dexamethasone and a methanol solvent;

finally, precisely measuring 1ml of each of the dexamethasone phosphate solution and the dexamethasone solution, placing the dexamethasone phosphate solution and the dexamethasone solution into a 100ml volumetric flask, adding the mobile phase to scale, and shaking up to obtain a system applicability solution containing 10 mu g of dexamethasone phosphate and 10 mu g of dexamethasone per 1 ml;

(2) preparation of a test solution: taking 0.2ml of dexamethasone phosphate esterification reaction liquid, adding 0.1ml of concentrated hydrochloric acid and 1ml of water, shaking up, adding a mobile phase to dilute to 10ml, and mixing uniformly to obtain a test solution;

(3) the blank solution is 10ml of mobile phase;

(4) detecting the cefazedone phosphate in the test sample solution by adopting a high performance liquid chromatography;

the preparation process of the mobile phase comprises the following steps: 45ml of triethylamine solution, 47.5ml of methanol and 7.5ml of acetonitrile are mixed uniformly to obtain a mobile phase.

2. The method for detecting dexamethasone in the reaction liquid in the esterification stage in the production process of dexamethasone sodium phosphate as claimed in claim 1, wherein the method comprises the following steps: in the step (1), the specific preparation process of the dexamethasone phosphate stock solution comprises the following steps: precisely weighing 25mg of dexamethasone phosphate reference substance, placing the dexamethasone phosphate reference substance in a 25ml volumetric flask, adding methanol for dissolving, fixing the volume to a scale, and shaking up to serve as dexamethasone phosphate stock solution;

the preparation process of the dexamethasone stock solution comprises the following steps: precisely weighing 25mg of dexamethasone, placing the dexamethasone into a 25ml volumetric flask, adding methanol for dissolving, fixing the volume to a scale, and shaking up to serve as dexamethasone stock solution.

3. The method for detecting dexamethasone in the reaction liquid in the esterification stage in the production process of dexamethasone sodium phosphate as claimed in claim 1, wherein the method comprises the following steps: in the step (4), the chromatographic conditions of the high performance liquid chromatography are as follows:

a) flow rate: 1.0 ml/min;

b) detection wavelength: 242 nm;

c) column temperature: 30 ℃;

d) sample introduction amount: 20 mu l of the mixture;

e) packing and column length of chromatographic column: octadecylsilane chemically bonded silica, 250X 4.6 mm;

4. the method for detecting dexamethasone in the reaction liquid in the esterification stage in the production process of dexamethasone sodium phosphate as claimed in claim 1, wherein the method comprises the following steps: in the step (4), the preparation process of the triethylamine solution is as follows: 7.5ml of triethylamine is taken, water is added for dilution to 1000ml, and the pH value is adjusted to 3.0 +/-0.05 by phosphoric acid.

5. The method for detecting dexamethasone in the reaction liquid in the esterification stage in the production process of dexamethasone sodium phosphate as claimed in claim 1, wherein the method comprises the following steps: in the step (4), the sample injection sequence of the high performance liquid chromatography is as follows: i) a blank solution; ii) a system suitability solution; iii) a test solution.

6. The method for detecting dexamethasone in the reaction liquid in the esterification stage in the production process of dexamethasone sodium phosphate as claimed in claim 5, wherein the method comprises the following steps: in the step (4), the specific operation process of the high performance liquid chromatography is as follows: respectively taking 20 mu l of blank solution, 20 mu l of system applicability solution and 20 mu l of sample solution, sequentially injecting the blank solution, the system applicability solution and the sample solution into a high performance liquid chromatograph according to a sample injection sequence, recording a chromatogram until the retention time of a main component is 2 times, wherein in the system applicability solution chromatogram, the theoretical plate number is calculated according to a dexamethasone phosphate peak and is not less than 3000, and the separation degree of the dexamethasone phosphate and the dexamethasone phosphate peak is more than 4.4.

Technical Field

The invention belongs to the technical field of detection methods, and particularly relates to a method for detecting dexamethasone in reaction liquid in an esterification stage in a production process of dexamethasone sodium phosphate.

Background

In view of the above situation, we disclose a detection standard and a detection method for the dexamethasone phosphate reaction solution so as to better control the production process of dexamethasone sodium phosphate, thereby improving the product quality of the dexamethasone sodium phosphate.

Disclosure of Invention

The invention aims to provide a method for detecting dexamethasone in reaction liquid in an esterification stage in a production process of dexamethasone sodium phosphate. By detecting the dexamethasone phosphate in the reaction liquid in the esterification stage, the quality control condition of the dexamethasone sodium phosphate in the production process can be better mastered.

The realization process of the invention is as follows:

the method for detecting dexamethasone in reaction liquid in the esterification stage in the production process of dexamethasone sodium phosphate comprises the following steps:

(1) preparation of system suitability solution:

firstly, preparing a dexamethasone phosphate stock solution by using a dexamethasone phosphate reference substance and a methanol solvent;

then, preparing dexamethasone stock solution by using dexamethasone and a methanol solvent;

finally, precisely measuring 1ml of each of the dexamethasone phosphate solution and the dexamethasone solution, placing the dexamethasone phosphate solution and the dexamethasone solution into a 100ml volumetric flask, adding the mobile phase to scale, and shaking up to obtain a system applicability solution containing 10 mu g of dexamethasone phosphate and 10 mu g of dexamethasone per 1 ml;

(2) preparation of a test solution: taking 0.2ml of dexamethasone phosphate esterification reaction liquid, adding 0.1ml of concentrated hydrochloric acid and 1ml of water, shaking up, adding a mobile phase to dilute to 10ml, and mixing uniformly to obtain a test solution;

(3) the blank solution is 10ml of mobile phase;

(4) detecting the cefazedone phosphate in the test sample solution by adopting a high performance liquid chromatography;

the preparation process of the mobile phase comprises the following steps: 45ml of triethylamine solution, 47.5ml of methanol and 7.5ml of acetonitrile are mixed uniformly to obtain a mobile phase.

Further, in the step (1), the specific preparation process of the dexamethasone phosphate stock solution comprises the following steps: precisely weighing 25mg of dexamethasone phosphate reference substance, placing the dexamethasone phosphate reference substance in a 25ml volumetric flask, adding methanol for dissolving, fixing the volume to a scale, and shaking up to serve as dexamethasone phosphate stock solution;

the preparation process of the dexamethasone stock solution comprises the following steps: precisely weighing 25mg of dexamethasone, placing the dexamethasone into a 25ml volumetric flask, adding methanol for dissolving, fixing the volume to a scale, and shaking up to serve as dexamethasone stock solution.

Further, in the step (4), the chromatographic conditions of the high performance liquid chromatography are as follows:

a) flow rate: 1.0 ml/min;

b) detection wavelength: 242 nm;

c) column temperature: 30 ℃;

d) sample introduction amount: 20 mu l of the mixture;

e) packing and column length of chromatographic column: octadecylsilane chemically bonded silica, 250X 4.6 mm;

further, in the step (4), the preparation process of the triethylamine solution is as follows: 7.5ml of triethylamine is taken, water is added for dilution to 1000ml, and the pH value is adjusted to 3.0 +/-0.05 by phosphoric acid.

Further, in the step (4), the sample injection sequence of the high performance liquid chromatography is as follows: i) a blank solution; ii) a system suitability solution; iii) a test solution.

Further, in the step (4), the specific operation process of the high performance liquid chromatography is as follows: respectively taking 20 mu l of blank solution, 20 mu l of system applicability solution and 20 mu l of sample solution, sequentially injecting the blank solution, the system applicability solution and the sample solution into a high performance liquid chromatograph according to a sample injection sequence, recording a chromatogram until the retention time of a main component is 2 times, wherein in the system applicability solution chromatogram, the theoretical plate number is calculated according to a dexamethasone phosphate peak and is not less than 3000, and the separation degree of the dexamethasone phosphate and the dexamethasone phosphate peak is more than 4.4.

The invention has the advantages that: the method can detect whether the dexamethasone reacts completely in the esterification reaction of the dexamethasone sodium phosphate, control the content of dexamethasone serving as an impurity in the product and improve the yield.

Drawings

FIG. 1 is a chromatogram of a first run of a blank solution;

FIG. 2 is a graph of a peak table for a first run of a blank solution;

FIG. 3 is a chromatogram of a second run of a blank solution;

FIG. 4 is a graph of a peak table for a second run of a blank solution;

FIG. 5 is a chromatogram of a third run of a blank solution;

FIG. 6 is a peak chart of a third experiment of a blank solution;

FIG. 7 is a chromatogram of a first experiment of a system suitability solution;

FIG. 8 is a graph of a peak table for a first run of a system suitability solution;

FIG. 9 is a chromatogram of a second run of a system suitability solution;

FIG. 10 is a chart of peaks from a second run of a system suitability solution;

FIG. 11 is a chromatogram of a third run of a system suitability solution;

FIG. 12 is a chart of peaks from a third experiment of a system suitability solution;

FIG. 13 is a chromatogram of a first experiment of a test solution;

FIG. 14 is a graph of a peak profile for a first run of a test sample solution;

FIG. 15 is a chromatogram of a second experiment of a test solution;

FIG. 16 is a graph of a peak profile for a second experiment of a test sample solution;

FIG. 17 is a chromatogram of a third experiment of a test solution;

FIG. 18 is a graph of a peak profile for a third experiment of a test sample solution;

FIG. 19 is a schematic diagram of the reaction process for producing dexamethasone sodium phosphate from dexamethasone.

Detailed Description

The present invention will be further described with reference to the following examples.

The invention discloses a method for detecting dexamethasone phosphate reaction solution by controlling the middle of an esterification stage in a production process of dexamethasone sodium phosphate so as to better monitor the middle process, thereby improving the product quality of the dexamethasone sodium phosphate.

Example 1

The method for detecting dexamethasone in reaction liquid in the esterification stage in the production process of dexamethasone sodium phosphate comprises the following steps:

(one) dexamethasone residues: HPLC

(II) instruments and appliances: analytical balance, pipette, high performance liquid chromatograph.

(III) reagent and test solution: dexamethasone, dexamethasone phosphate control, acetonitrile, methanol, triethylamine, phosphoric acid, concentrated hydrochloric acid (assay pure).

(IV) solution preparation

(1) Preparation of system suitability solution:

firstly, the specific preparation process of the dexamethasone phosphate stock solution comprises the following steps: precisely weighing 25mg of dexamethasone phosphate reference substance, placing the dexamethasone phosphate reference substance in a 25ml volumetric flask, adding methanol for dissolving, fixing the volume to a scale, and shaking up to serve as dexamethasone phosphate stock solution;

then, the specific preparation process of the dexamethasone stock solution is as follows: precisely weighing 25mg of dexamethasone, placing the dexamethasone into a 25ml volumetric flask, adding methanol for dissolving, fixing the volume to a scale, and shaking up to serve as dexamethasone stock solution;

finally, precisely measuring 1ml of each of the dexamethasone phosphate solution and the dexamethasone solution, placing the dexamethasone phosphate solution and the dexamethasone solution into a 100ml volumetric flask, adding the mobile phase to scale, and shaking up to obtain a system applicability solution containing 10 mu g of dexamethasone phosphate and 10 mu g of dexamethasone per 1 ml;

(2) preparation of a test solution: taking 0.2ml of dexamethasone phosphate esterification reaction liquid, adding 0.1ml of concentrated hydrochloric acid and 1ml of water, shaking up, adding a mobile phase to dilute to 10ml, and mixing uniformly to obtain a test solution;

(3) the blank solution is 10ml of mobile phase;

(V) chromatographic conditions and sequence arrangement

1. Flow rate: 1.0ml/min

2. Detection wavelength: 242nm

3. Column temperature: 30 deg.C

4. Sample introduction amount: 20 μ l

5. Packing and column length of chromatographic column: octadecylsilane chemically bonded silica, 250X 4.6mm

6. Mobile phase: 45ml of triethylamine solution, 47.5ml of methanol and 7.5ml of acetonitrile are uniformly mixed to obtain a mobile phase; the preparation process of the triethylamine solution comprises the following steps: 7.5ml of triethylamine is taken, water is added for dilution to 1000ml, and the pH value is adjusted to 3.0 +/-0.05 by phosphoric acid.

7. The sample injection sequence of the high performance liquid chromatography is as follows: i) a blank solution; ii) a system suitability solution; iii) a test solution.

8. The specific operation process of the high performance liquid chromatography is as follows: respectively taking 20 mu l of blank solution, 20 mu l of system applicability solution and 20 mu l of sample solution, sequentially injecting the blank solution, the system applicability solution and the sample solution into a high performance liquid chromatograph according to a sample injection sequence, recording a chromatogram until the retention time of a main component is 2 times, wherein in the system applicability solution chromatogram, the theoretical plate number is calculated according to a dexamethasone phosphate peak and is not less than 3000, and the separation degree of the dexamethasone phosphate and the dexamethasone phosphate peak is more than 4.4.

9. Sample injection sequence editing and result:

TABLE 1 sample introduction sequence editing and results

Note that: the blank solution is used in the sequence to eliminate test deviation caused by instrument and reagent contamination, and the sequence is the test sequence of a batch of samples, and three batches of measurement are carried out. If the measurement of multiple batches is carried out simultaneously, the blank sample injection times and the reference sample injection times are increased properly.

With reference to table 1 and fig. 1 to 18, it can be seen whether dexamethasone completely reacts or remains in the reaction solution during the esterification stage in the production process of dexamethasone sodium phosphate. As can be seen from FIGS. 7-12, the retention time of dexamethasone phosphate in the system suitability solution is about 15min, with an area% of 41, and the retention time of dexamethasone is about 19-20min, with an area% of 58. As can be seen from FIGS. 13-18, the retention time of dexamethasone phosphate was 15min at about 96 area%, the retention time of dexamethasone was about 19-20min at about 1 area%, indicating that the dexamethasone was substantially reacted completely with less residue.

Acceptance criteria: if a chromatographic peak corresponding to the dexamethasone peak-out time appears in the chromatogram of the test solution, the area percent is not more than 5.0 percent according to the area normalization method.

After the method is adopted, the dexamethasone phosphate in the middle process of the production of the dexamethasone sodium phosphate product is controlled, whether the dexamethasone reacts completely in the material dexamethasone in the esterification reaction of the dexamethasone sodium phosphate is controlled, the content of the impurity dexamethasone in the product is controlled, and the yield of the dexamethasone sodium phosphate product is improved.

The foregoing is a more detailed description of the invention in connection with specific preferred embodiments and is not intended to limit the invention to the particular forms disclosed. For those skilled in the art to which the invention pertains, several simple deductions or substitutions can be made without departing from the spirit of the invention, and all shall be considered as belonging to the protection scope of the invention.