CN116087355A - Method for measuring content of diethyl phosphate in raw material medicine of present vitamin mod - Google Patents
Method for measuring content of diethyl phosphate in raw material medicine of present vitamin mod Download PDFInfo
- Publication number
- CN116087355A CN116087355A CN202211596332.1A CN202211596332A CN116087355A CN 116087355 A CN116087355 A CN 116087355A CN 202211596332 A CN202211596332 A CN 202211596332A CN 116087355 A CN116087355 A CN 116087355A
- Authority
- CN
- China
- Prior art keywords
- diethyl phosphate
- solution
- mobile phase
- reference substance
- taking
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/60—Construction of the column
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8624—Detection of slopes or peaks; baseline correction
- G01N30/8631—Peaks
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention relates to the technical field of medicine detection, in particular to a method for measuring the content of diethyl phosphate in a raw material medicine of the present vitamin mod, which comprises the following specific steps: taking a proper amount of diethyl phosphate reference substance, and preparing a diethyl phosphate solution with proper concentration by taking acetonitrile as a diluent to serve as a reference substance solution; taking a proper amount of the raw material medicine of the vitamin and taking acetonitrile as a solvent to prepare a sample solution; preparing a series of diethyl phosphate reference substance solutions, detecting by an LC-MS method, respectively obtaining peak areas of diethyl phosphate in the reference substance solutions, and performing standard curve fitting with the solution concentration; detecting the sample solution by an LC-MS method to obtain the peak area of the sample solution, and calculating the content of diethyl phosphate in the sample solution by an external standard curve method. The invention adopts the LC-MS method to detect the content of the diethyl phosphate in the raw material medicine of the present vitamin mod, and the measuring method has the characteristics of convenient operation, short detection time, high specificity and high precision and accuracy.
Description
Technical Field
The invention relates to the technical field of medicine detection, in particular to a method for measuring the content of diethyl phosphate in a raw material medicine of the present vitamin mod.
Background
The vitamin D (Tapinarof) is a medicine for treating adult plaque psoriasis, and a small amount of diethyl phosphate remains in the raw material medicine. Diethyl phosphate is often used as an extractant, a solvent and a polymerization catalyst, and has certain irritation, so that eyes and respiratory systems can be stimulated, and skin contact can be sensitized, so that the residual test of the diethyl phosphate is particularly important, and the determination method of the content of the diethyl phosphate is mainly a traditional titration method and an ion chromatography method at present.
The traditional titration method has low sensitivity and cannot meet the requirement of the amount of the residual diethyl phosphate in the raw material drug of the present vitamin mod; the specificity of the ion chromatography is poor, the qualitative detection of the diethyl phosphate cannot be carried out, the measurement time is too long, the chromatographic column and the consumable material cost are relatively expensive, and the detection cost is high.
Disclosure of Invention
The invention aims to provide a method for measuring the content of diethyl phosphate in a raw material drug of the present vitamin mod, so as to solve the problems in the prior art.
The invention is realized by the following technical scheme:
the method for determining the content of diethyl phosphate in the valmod bulk drug specifically comprises the following steps:
s1, taking a proper amount of diethyl phosphate reference substance, and preparing a diethyl phosphate solution with proper concentration by taking acetonitrile as a diluent to serve as a reference substance solution;
s2, taking a proper amount of the valmod raw material medicine, and preparing a sample solution by taking acetonitrile as a solvent;
s3, preparing a series of diethyl phosphate reference substance solutions, detecting by an LC-MS method, respectively obtaining peak areas of diethyl phosphate in the reference substance solutions, and performing standard curve fitting with the solution concentration;
s4, detecting the sample solution by an LC-MS method to obtain the peak area of the sample solution, and calculating the content of diethyl phosphate in the sample solution by an external standard curve method;
wherein, the chromatographic conditions of the LC-MS method in the steps S3 and S4 are as follows:
the chromatographic column adopts an ACE Excel 3C18 chromatographic column with the specification of 4.6X105 mm and 3 mu m, the column temperature is 30 ℃, the sample injection amount is 5 mu L, and the flow rate is 0.6mL/min; the temperature of the sample feeding disc is room temperature; the detector is a mass spectrum detector, the ion source is ESI, and the positive ion mode is adopted.
As a further scheme of the invention, in the steps S3 and S4, 0.1% formic acid water solution is taken as a mobile phase A, acetonitrile is taken as a mobile phase B, and the elution conditions are as follows: the gradient elution mode is adopted, and the gradient elution program is as follows: the volume percentage of the mobile phase B is 5% within 0-3 min; 3-6 min, the volume percentage of the mobile phase B is changed from 5% to 90%; the volume percentage of the mobile phase B is 95% after 6-8 min; the volume percentage of the mobile phase B is changed from 90% to 0 after 8-8.1 min; 8.1-12 min, and the volume percentage of the mobile phase B is 5%.
The specific gradient elution procedure is as follows:
| T/min | 0.1% formic acid | Acetonitrile | |
| 0 | 95％ | 5％ | |
| 3 | 95％ | 5％ | |
| 6 | 10％ | 90％ | |
| 8 | 10％ | 90％ | |
| 8.1 | 95％ | 5％ | |
| 12 | 95％ | 5％ |
As a further aspect of the present invention, in step S3, the concentration of the series of diethyl phosphate control solutions was 0.01. Mu.g/mL, 0.05. Mu.g/mL, 0.10. Mu.g/mL, 0.15. Mu.g/mL, and 0.20. Mu.g/mL, respectively.
Compared with the prior art, the invention has the beneficial effects that:
the invention adopts the liquid chromatograph-mass spectrometer to detect the content of the diethyl phosphate in the raw material medicine of the vitamin mod, and the measuring method has the characteristics of convenient operation, short detection time, high specificity and high precision and accuracy.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed for the description of the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic diagram of a standard curve of a diethyl phosphate control solution of the present invention;
FIG. 2 is a graph of a blank solution of the present invention;
FIG. 3 is a graph showing the detection of a sample solution of the present invention;
FIG. 4 is a graph showing the detection of a control solution of the present invention;
FIG. 5 is a graph showing the detection of 100% test sample labeling solution according to the present invention.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1:
the embodiment provides a method for measuring the content of diethyl phosphate in a raw material drug of the present vitamin mod, which specifically comprises the following steps:
s1, taking a proper amount of diethyl phosphate reference substance, and preparing a diethyl phosphate solution with proper concentration by taking acetonitrile as a diluent to serve as a reference substance solution;
s2, taking a proper amount of the valmod raw material medicine, and preparing a sample solution by taking acetonitrile as a solvent;
s3, preparing reference substance solutions of diethyl phosphate with the concentration of 0.01 mug/mL, 0.05 mug/mL, 0.10 mug/mL, 0.15 mug/mL and 0.20 mug/mL respectively, detecting by an LC-MS method, respectively obtaining peak areas of diethyl phosphate in the reference substance solutions, specifically data are shown in table 1, performing standard curve fitting with the concentration of the solutions, wherein the obtained standard curve formula is y= 1969849.0425x-12622.5533, R 2 = 0.9955, the fitted standard curve is shown in fig. 1, and the result shows that diethyl phosphate exhibits good linear relationship in the concentration range;
table 1:
s4, detecting the sample solution by an LC-MS method to obtain the peak area of the sample solution, and calculating the content of diethyl phosphate in the sample solution by an external standard curve method;
wherein, the chromatographic conditions of the LC-MS method in the steps S3 and S4 are as follows:
the chromatographic column adopts an ACE Excel 3C18 chromatographic column with the specification of 4.6X105 mm and 3 mu m, the column temperature is 30 ℃, the sample injection amount is 5 mu L, and the flow rate is 0.6mL/min; the temperature of the sample feeding disc is room temperature; the detector is a mass spectrum detector, the ion source is ESI, and the positive ion mode is adopted.
In the steps S3 and S4, 0.1% formic acid aqueous solution is taken as a mobile phase A, acetonitrile is taken as a mobile phase B, and the elution conditions are as follows: the gradient elution mode is adopted, and the gradient elution program is as follows: the volume percentage of the mobile phase B is 5% within 0-3 min; 3-6 min, the volume percentage of the mobile phase B is changed from 5% to 90%; the volume percentage of the mobile phase B is 95% after 6-8 min; the volume percentage of the mobile phase B is changed from 90% to 0 after 8-8.1 min; 8.1-12 min, and the volume percentage of the mobile phase B is 5%. The specific gradient elution procedure is shown in table 2.
Table 2:
| T/min | 0.1% formic acid | Acetonitrile | |
| 0 | 95％ | 5％ | |
| 3 | 95％ | 5％ | |
| 6 | 10％ | 90％ | |
| 8 | 10％ | 90％ | |
| 8.1 | 95％ | 5％ | |
| 12 | 95％ | 5％ |
And detecting blank solution, test sample solution, reference substance solution and 100% test sample and reference substance solution (test sample and reference solution) according to the chromatographic conditions and operation steps, wherein the detection results are shown in figures 2-5. The detection results of the attached figures 2-5 show that the blank solution and the sample solution are free from interference, and the sample adding standard solution has obvious response, so that the detection method provided by the invention has high specificity and can be used for qualitatively detecting the diethyl phosphate.
An appropriate amount of control solution was added to the test solution for accuracy experiments, and the accuracy results are shown in table 3.
Table 3:
| sequence number | Peak area | Addition (ng) | Measured (ng) | Original quantity (ng) | Recovery rate |
| 50％-1 | 99035 | 4.6661 | 5.4340 | 0 | 116％ |
| 50％-2 | 97003 | 4.6661 | 5.3225 | 0 | 114％ |
| 50％-3 | 95191 | 4.6661 | 5.2231 | 0 | 112％ |
| 100％-1 | 189866 | 9.3322 | 10.4179 | 0 | 112％ |
| 100％-2 | 187229 | 9.3322 | 10.2732 | 0 | 110％ |
| 100％-3 | 188372 | 9.3322 | 10.3359 | 0 | 111％ |
| 150％-4 | 278729 | 13.9982 | 15.2938 | 0 | 109％ |
| 150％-2 | 281726 | 13.9982 | 15.4583 | 0 | 110％ |
| 150％-3 | 275487 | 13.9982 | 15.1159 | 0 | 108％ |
As shown in the table, the content of diethyl phosphate in the raw material medicine of the present vitamin mod is detected by adopting a liquid chromatograph-mass spectrometer, and the detection result has high precision and accuracy and good repeatability.
The preferred embodiments of the invention disclosed above are intended only to assist in the explanation of the invention. The preferred embodiments are not exhaustive or to limit the invention to the precise form disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, to thereby enable others skilled in the art to best understand and utilize the invention. The invention is limited only by the claims and the full scope and equivalents thereof.
Claims (3)
1. The method for determining the content of diethyl phosphate in the valmod bulk drug is characterized by comprising the following steps of:
s1, taking a proper amount of diethyl phosphate reference substance, and preparing a diethyl phosphate solution with proper concentration by taking acetonitrile as a diluent to serve as a reference substance solution;
s2, taking a proper amount of the valmod raw material medicine, and preparing a sample solution by taking acetonitrile as a solvent;
s3, preparing a series of diethyl phosphate reference substance solutions, detecting by an LC-MS method, respectively obtaining peak areas of diethyl phosphate in the reference substance solutions, and performing standard curve fitting with the solution concentration;
s4, detecting the sample solution by an LC-MS method to obtain the peak area of the sample solution, and calculating the content of diethyl phosphate in the sample solution by an external standard curve method;
wherein, the chromatographic conditions of the LC-MS method in the steps S3 and S4 are as follows:
the chromatographic column adopts an ACE Excel 3C18 chromatographic column with the specification of 4.6X105 mm and 3 mu m, the column temperature is 30 ℃, the sample injection amount is 5 mu L, and the flow rate is 0.6mL/min; the temperature of the sample feeding disc is room temperature; the detector is a mass spectrum detector, the ion source is ESI, the positive ion mode, and the ion is 155.
2. The method for determining the content of diethyl phosphate in a crude drug of present vmod according to claim 1, wherein in steps S3 and S4, 0.1% formic acid aqueous solution is used as mobile phase a, acetonitrile is used as mobile phase B, and elution conditions are: the gradient elution mode is adopted, and the gradient elution program is as follows: the volume percentage of the mobile phase B is 5% within 0-3 min; 3-6 min, the volume percentage of the mobile phase B is changed from 5% to 90%; the volume percentage of the mobile phase B is 95% after 6-8 min; the volume percentage of the mobile phase B is changed from 90% to 0 after 8-8.1 min; 8.1-12 min, and the volume percentage of the mobile phase B is 5%.
3. The method according to any one of claims 1-2, wherein in step S3, the concentration of the series of diethyl phosphate control solutions is 0.01 μg/mL, 0.05 μg/mL, 0.10 μg/mL, 0.15 μg/mL and 0.20 μg/mL, respectively.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211596332.1A CN116087355A (en) | 2022-12-12 | 2022-12-12 | Method for measuring content of diethyl phosphate in raw material medicine of present vitamin mod |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211596332.1A CN116087355A (en) | 2022-12-12 | 2022-12-12 | Method for measuring content of diethyl phosphate in raw material medicine of present vitamin mod |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116087355A true CN116087355A (en) | 2023-05-09 |
Family
ID=86200198
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211596332.1A Withdrawn CN116087355A (en) | 2022-12-12 | 2022-12-12 | Method for measuring content of diethyl phosphate in raw material medicine of present vitamin mod |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116087355A (en) |
-
2022
- 2022-12-12 CN CN202211596332.1A patent/CN116087355A/en not_active Withdrawn
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113671077B (en) | Method for detecting acryloyl chloride and related substances | |
| CN107870209B (en) | Method for determining impurity content in linagliptin bulk drug | |
| CN113295805B (en) | Method for detecting hydrazine hydrate in medicine | |
| CN116087355A (en) | Method for measuring content of diethyl phosphate in raw material medicine of present vitamin mod | |
| CN111208215B (en) | Method for detecting impurity 2-mercaptobenzothiazole in ceftriaxone sodium | |
| CN110618210A (en) | Method for detecting content of anions in motherwort injection | |
| CN112526028B (en) | Method for determining content of dithiothreitol in protein solution | |
| CN108760937B (en) | Determination of residual ethylenediamine in caspofungin acetate and application thereof | |
| CN111337620B (en) | Method for detecting content of 3-amino-2-piperidone in compound amino acid injection | |
| CN114137120A (en) | Method for detecting related substances in rapamycin drug stent | |
| CN112763591A (en) | Method for determining penicillin content in protein solution | |
| CN114047279A (en) | Method for measuring residual quantity of N-methylpiperazine in drug intermediate and bulk drug by high performance liquid chromatography-mass spectrometry | |
| CN108572223B (en) | Method for determining activity inducing substance in polypeptide | |
| CN112710762A (en) | Method for measuring residual quantity of dimethyl sulfate | |
| CN108008035B (en) | Method for detecting purity of 3-ethoxy-4-methoxybenzaldehyde | |
| CN114200067B (en) | High performance liquid chromatography analysis method for 6-bromo-3-hydroxy pyrazine-2-carboxamide and impurities | |
| CN116399984B (en) | Method for measuring residual quantity of tetrabutylammonium iodide in WXTJ0262 bulk drug by utilizing liquid phase-mass spectrum combined method | |
| CN115951000B (en) | Method for detecting oxalate in calcium gluconate | |
| CN114235972B (en) | Method for determining content of linagliptin impurity RBP-1 | |
| CN113777204B (en) | Detection method of p-hydroxyacetophenone related substances | |
| CN113884604B (en) | Method for determining content of cellulose in auxiliary material in medicine by liquid phase method | |
| CN114646700B (en) | Detection method of (S) -pyrrolidine-2-formonitrile hydrochloride | |
| CN114047280B (en) | Method for detecting content of non-volatile unknown substances in medical instrument leaching liquor | |
| CN111103374B (en) | Method for measuring content of 2, 6-tetramethylpiperidine oxide in cinacalcet hydrochloride | |
| CN107677742B (en) | Quantitative detection method for residual hydroxyethyl acrylate in polymer synthesis process |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20230509 |
|
| WW01 | Invention patent application withdrawn after publication |