CN111678998B - Separation and detection method for enantiomers in raltitrexed - Google Patents
Separation and detection method for enantiomers in raltitrexed Download PDFInfo
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- CN111678998B CN111678998B CN202010362885.5A CN202010362885A CN111678998B CN 111678998 B CN111678998 B CN 111678998B CN 202010362885 A CN202010362885 A CN 202010362885A CN 111678998 B CN111678998 B CN 111678998B
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- G—PHYSICS
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- 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
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- 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
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- G—PHYSICS
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- 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
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- 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
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Abstract
The invention discloses a separation and detection method of enantiomers in raltitrexed. The method is a high performance liquid chromatography, and the selected stationary phase is an octadecyl bonded silica gel column; the mobile phase is acetonitrile and cyclodextrin water solution or cyclodextrin derivative water solution. The method is mature, universal, simple and quick, is suitable for separating and detecting the enantiomers in raltitrexed and preparations, and has the characteristics of good separation degree, high sensitivity, strong specificity and the like.
Description
Technical Field
The invention relates to the field of drug analysis, in particular to a separation and detection method of enantiomers in raltitrexed.
Background
Raltitrexed for injection was co-developed by the Zeneca pharmaceutical and tumour research centre (UK), marketed in the UK 3 months 1996 under the trade name Tomudex for the treatment of mid-late colorectal cancer, and was thereafter introduced in more than 40 other countries, including the european union, eastern europe and south america in most countries. The indications are as follows: when the patients cannot receive the combined chemotherapy, the product can be singly used for treating the patients with advanced colorectal cancer which is not suitable for 5-FU/calcium folinate. Compared with the conventional first-line drug 5-fluorouracil for treating colorectal cancer, the raltitrexed has stronger inhibiting effect on a colon cancer cell line and lower toxicity; more convenient use, can obviously improve the life quality of patients and the like. Raltitrexed, as the first new generation of water-soluble thymidylate synthase inhibitor for treating colorectal cancer in the last forty years, is a first-line cytotoxic chemotherapy drug and can become an effective substitute drug for treating advanced colorectal cancer by taking 5-fluorouracil as a basic scheme. The molecular formula is C21H22N4O6And S has the chemical structural formula shown in the specification.
Certain enantiomers of raltitrexed exist, and the chemical structural formula is shown below.
The enantiomer has no drug effect and affects the quality of the medicine. The raltitrexed for injection is freeze-dried powder injection for injection, the specification is 2mg, and the prescription of unit dose comprises the following components:
| composition (I) | Dosage of |
| Raltitrexed | 2mg |
| Mannitol | 203mg |
| Anhydrous disodium hydrogen phosphate | 0.8mg |
| Sodium hydroxide | 0-1mg |
| Water for injection | Adding to 2ml |
The method comprises the steps of dissolving raltitrexed in water for injection with disodium hydrogen phosphate as an aid, adjusting the pH value to 7.2-7.6 with sodium hydroxide, and enabling the raltitrexed preparation to exist in a sodium salt form. Therefore, the enantiomeric impurities in raltitrexed starting materials and formulations need to be quality controlled. The method controls the content of the enantiomers in the raw materials and the preparations of the raltitrexed, and has great significance for improving the quality of the medicine and ensuring the medication safety of patients.
However, the separation and detection of enantiomers is always the key point and difficulty of quality control in drug synthesis, especially raltitrexed contains an aminoglutaric acid structure, the raw material drug exists in a carboxylic acid form, the preparation of the raltitrexed exists in a sodium salt form, and the difference between the two properties is large; the raw materials in the unit dose preparation of the raltitrexed account for 2mg and the mannitol accounts for 203mg, thus increasing the difficulty of separating the enantiomers of the raltitrexed preparation.
Disclosure of Invention
The following is a summary of the subject matter described in detail herein. This summary is not intended to limit the scope of the invention.
The invention develops a method for separating and detecting enantiomers in a raltitrexed bulk drug and a preparation thereof by reverse phase chromatography. The following problems are solved:
chinese patent application CN101221147A discloses a method for detecting an enantiomer of raltitrexed by capillary electrophoresis. Electroosmosis of the capillary electrophoresis method is changed due to sample composition, and the reproducibility is poor; the capillary has small diameter, and when ultraviolet absorption spectroscopy is used, the sensitivity is low, and quantitative detection has certain difficulty.
Chinese patent application CN107167535A discloses a method for detecting raltitrexed enantiomer by reversed phase liquid chromatography. In the method, triethanolamine is added into a mobile phase, the triethanolamine has the properties of tertiary amine and alcohol, and raltitrexed has a glutaric acid structure and can form ester with the triethanolamine, so that the solution stability of the method is poor.
Chinese patent application CN109283279A discloses a method for separating and determining raltitrexed and its enantiomer by high performance liquid chromatography. The active ingredient raltitrexed in the raltitrexed preparation exists in a disodium salt form, the skeleton agent is mannitol, so the raltitrexed preparation can be dissolved only by water phase, and the method adopts a normal phase liquid chromatography system, can not use water phase, can only carry out isomer detection on the raltitrexed raw material medicine, can not be compatible with the raltitrexed preparation, and has narrow application range.
The invention provides a separation and detection method of an enantiomer in a raltitrexed bulk drug and a preparation thereof. The method is based on reversed phase liquid chromatography, is mature, universal, simple and quick, and is suitable for separating and detecting raltitrexed and chiral isomers thereof. The method has the characteristics of good separation degree, high sensitivity, strong specificity and the like, and overcomes the defects in the prior art.
The invention provides a separation and detection method of enantiomers in raltitrexed, which comprises the following steps:
1) adopting high performance liquid chromatography, setting the chromatographic conditions as follows:
a chromatographic column: octadecylsilane chemically bonded silica gel column is used as a stationary phase;
detection wavelength: 224nm to 230 nm;
column temperature: 3-42 ℃;
sample introduction amount: 1-20 μ l;
flow rate: 0.8ml/min-1.2 ml/min;
mobile phase: the composite material consists of a mobile phase A and a mobile phase B, wherein the mobile phase A is an aqueous solution of cyclodextrin or an aqueous solution of a cyclodextrin derivative, and the pH value is adjusted to 3.6-6.0 by phosphoric acid; the mobile phase B is acetonitrile; the volume ratio of the mobile phase A to the mobile phase B is (600- & ltSUB & gt 1000) & lt/SUB & gt (60-200); and carrying out isocratic elution or gradient elution;
2) preparing a test solution:
taking a proper amount of the raltitrexed raw material medicine or the raltitrexed preparation, adding sodium hydroxide solution to dissolve the raw material medicine or the raltitrexed preparation, and adding water to dilute the raw material medicine or the raltitrexed preparation to prepare a raltitrexed test solution;
3) measuring by high performance liquid chromatography under the chromatographic condition of step 1), precisely measuring the sample solution, injecting into a liquid chromatograph, and recording the chromatogram.
In the embodiment of the invention, the raltitrexed preparation is raltitrexed for injection, and is a freeze-dried powder injection for injection.
In an embodiment of the invention, optionally, the concentration of the sodium hydroxide solution is 0.1 mol/L.
In an embodiment of the invention, the aqueous solution of cyclodextrin has a cyclodextrin that is alpha-cyclodextrin, beta-cyclodextrin, or gamma-cyclodextrin; the cyclodextrin derivative in the aqueous solution of the cyclodextrin derivative is propyl-beta-cyclodextrin, sulfonated-beta-cyclodextrin, or carboxymethyl-beta-cyclodextrin.
In the above embodiment, the concentration of the cyclodextrin in the aqueous solution of cyclodextrin or the cyclodextrin derivative in the aqueous solution of cyclodextrin derivative is 0.05% to 2.0% g/ml.
In the above embodiments, the mobile phase a is an aqueous solution of sulfonated- β -cyclodextrin at a concentration ranging from 0.05% to 2.0% g/ml, preferably from 0.6% to 1.2% g/ml, more preferably 0.8%, 0.9% g/ml or 1.0% g/ml.
In the above embodiment, the mobile phase a is adjusted to a pH of 3.0 to 6.0, preferably 5.0, with phosphoric acid.
In the above embodiment, the volume ratio of the mobile phase A to the mobile phase B is preferably (910-930) (70-90), and more preferably 920: 80.
In the above embodiment, preferably, the detection wavelength is 226 nm.
In the above embodiment, preferably, the column temperature is 40 ℃.
In the above embodiment, preferably, the sample amount is 10. mu.l.
In the above embodiment, preferably, the flow rate is 1.0 ml/min.
In the above embodiment, the concentration of the test solution is 0.5mg/ml to 2mg/ml, preferably 0.5 mg/ml.
In the above embodiment, the step 2) further comprises preparing a control solution: precisely measuring 1ml of the test solution, placing the test solution in a 200ml measuring flask, diluting the test solution to a scale with water, and shaking up to obtain a control solution.
In the above embodiment, the step 2) further comprises preparing a resolution solution: taking about 10mg of raltitrexed reference substance, precisely weighing, placing in a 20ml measuring flask, precisely weighing 1ml of raltitrexed enantiomer stock solution, adding 2ml of 0.1mol/L sodium hydroxide solution for dissolving, adding water for diluting to a scale, shaking up, and taking as a resolution solution; wherein, the preparation of the raltitrexed enantiomer stock solution comprises the following steps: about 25mg of raltitrexed enantiomer is taken, precisely weighed, placed in a 500ml measuring flask, dissolved and diluted to the scale by adding 0.1mol/L sodium hydroxide solution, and shaken up to be used as optical isomer stock solution.
Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. The objectives and other advantages of the invention may be realized and attained by the structure particularly pointed out in the written description and drawings.
Drawings
The accompanying drawings are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the example serve to explain the principles of the invention and not to limit the invention.
FIG. 1 is a high performance liquid chromatogram of a test solution in example 1 of the present invention;
FIG. 2 is a high performance liquid chromatogram of a test solution in example 2 of the present invention;
FIG. 3 is a high performance liquid chromatogram of the resolution solution in example 3 of the present invention;
FIG. 4 is a high performance liquid chromatogram of a resolution solution of a comparative example of the present invention;
FIG. 5 is a high performance liquid chromatogram of a test solution in a comparative example of the present invention;
FIG. 6 is a high performance liquid chromatogram of an isomer solution in a comparative example of the present invention;
FIG. 7 is a high performance liquid chromatogram of an aqueous solution of sulfonated-beta-cyclodextrin having triethanolamine changed to 0.8% g/ml in a comparative example of the present invention;
FIG. 8 is a high performance liquid chromatogram of the optimized ratio of 0.8% g/ml aqueous sulfonated-beta-cyclodextrin solution to organic phase in the mobile phase of a comparative example of the present invention.
Detailed Description
Hereinafter, embodiments of the present invention will be described in detail in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be noted that the embodiments and features of the embodiments in the present application may be arbitrarily combined with each other without conflict.
Example 1
Detection of enantiomers of raltitrexed bulk drug
1.1 chromatographic conditions
The instrument comprises the following steps: agilent 1260 high performance liquid chromatograph;
a chromatographic column: welch Ultimate XB-C18, 3.9X 150mm, 5 μm;
mobile phase: adding 1000ml of water into 8g of sulfonated beta-cyclodextrin serving as a mobile phase A for dissolving, and adding phosphoric acid for regulating the pH value to 5.0; the mobile phase B is acetonitrile, and the volume ratio of the mobile phase A to the mobile phase B is 920: 80;
detection wavelength: 226 nm;
column temperature: 40 ℃;
sample introduction amount: 10 mu l of the mixture;
flow rate: 1.0 ml/min;
operating time: and (3) 30 min.
1.2 sample preparation
Blank solution: 0.1mol/L sodium hydroxide solution.
Test solution: taking 10mg of raltitrexed raw material medicine, adding 2ml of 0.1mol/L sodium hydroxide solution for dissolving, adding water for diluting to a 20ml measuring flask, and shaking up to be used as a test solution.
Control solution: precisely measuring 1ml of the test solution, placing the test solution in a 200ml measuring flask, adding water to dilute the test solution to a scale, and shaking the test solution uniformly to serve as a control solution.
Raltitrexed enantiomer stock solutions: about 25mg of raltitrexed enantiomer is taken, precisely weighed, placed in a 500ml measuring flask, dissolved and diluted to the scale by adding 0.1mol/L sodium hydroxide solution, and shaken up to be used as optical isomer stock solution.
Enantiomeric positioning solution: precisely measuring 5ml of optical isomer stock solution, placing the optical isomer stock solution into a 50ml measuring flask, diluting the optical isomer stock solution to a scale with water, and shaking up to obtain the enantiomer positioning solution.
Resolution solution: taking about 10mg of raltitrexed reference substance, precisely weighing, placing in a 20ml measuring flask, precisely weighing 1ml of raltitrexed enantiomer stock solution, adding 2ml of 0.1mol/L sodium hydroxide solution for dissolving, adding water for diluting to a scale, shaking up, and taking as a resolution solution.
1.3 determination of
Injecting 10 μ l of the separation degree solution into a high performance liquid chromatograph, and recording a chromatogram, wherein the separation degree should be more than 1.5.
And respectively injecting 10 mu l of the test solution and 10 mu l of the control solution into a high performance liquid chromatograph, and recording the chromatogram. The enantiomeric contents were calculated according to the self-control method.
The high performance liquid chromatography results of the test solution are shown in figure 1, and it can be seen that raltitrexed and its enantiomer are completely separated under the conditions.
Example 2
Raltitrexed formulation enantiomer detection
The raltitrexed preparation is raltitrexed for injection, and the specification is as follows: 2mg, source: yangzhijiang pharmaceutical group, Inc.
2.1 chromatographic conditions
The instrument comprises the following steps: agilent 1260 high performance liquid chromatograph;
a chromatographic column: welch Ultimate XB-C18, 3.9X 150mm, 5 μm;
mobile phase: adding 1000ml of water into 8g of sulfonated beta-cyclodextrin serving as a mobile phase A for dissolving, and adding phosphoric acid for regulating the pH value to 5.0; the mobile phase B is acetonitrile, and the volume ratio of the mobile phase A to the mobile phase B is 920: 80;
detection wavelength: 226 nm;
column temperature: 40 ℃;
sample introduction amount: 10 mu l of the mixture;
flow rate: 1.0 ml/min;
operating time: and (3) 30 min.
2.2 sample preparation
Blank solution: 0.1mol/L sodium hydroxide solution.
Test solution: taking 5 raltitrexed samples for injection, adding 2ml of 0.1mol/L sodium hydroxide solution for dissolving, adding water for diluting to a 20ml measuring flask, and shaking up to be used as a test solution.
Control solution: precisely measuring 1ml of the test solution, placing the test solution in a 200ml measuring flask, diluting the test solution to a scale with water, and shaking up to obtain a control solution.
Raltitrexed enantiomer stock solutions: about 25mg of raltitrexed enantiomer is taken, precisely weighed, placed in a 500ml measuring flask, dissolved and diluted to the scale by adding 0.1mol/L sodium hydroxide solution, and shaken up to be used as optical isomer stock solution.
Resolution solution: taking about 10mg of raltitrexed reference substance, precisely weighing, placing in a 20ml measuring flask, precisely weighing 1ml of raltitrexed enantiomer stock solution, adding 2ml of 0.1mol/L sodium hydroxide solution for dissolving, adding water for diluting to a scale, shaking up, and taking as a resolution solution.
2.3 determination of
And (5) injecting 10 mu l of the resolution solution into a high performance liquid chromatograph, and recording the chromatogram. The degree of separation should be greater than 1.5.
And respectively injecting 10 mu l of the test solution and 10 mu l of the control solution into a high performance liquid chromatograph, and recording the chromatogram. The enantiomeric contents were calculated according to the self-control method.
The HPLC result of the test solution is shown in FIG. 2, which shows that the raltitrexed preparation and its enantiomer are completely separated under the condition.
Example 3
Durability of Raltitrexed enantiomer process
The instrument comprises the following steps: agilent 1260 high performance liquid chromatograph;
the tolerance of the detection method was examined when the chromatographic conditions were varied. The results of the detection under the varied chromatographic conditions were as follows:
the high performance liquid chromatography result of the resolution solution with the mobile phase pH value of 5.2 is shown in figure 3, and the complete separation of the enantiomers of the raltitrexed under the condition can be seen.
And (4) conclusion: under various variable chromatographic conditions, the resolution between the optical isomer and the main peak in each solution meets the requirement; compared with the normal condition, the ratio of the content of the optical isomer to the normal condition is 80.0-120.0%, and the method has good durability.
Comparative example
Raltitrexed preparation enantiomer detection by adopting CN107167535A liquid phase condition
1.1 chromatographic conditions
The instrument comprises the following steps: agilent 1260 high performance liquid chromatograph;
a chromatographic column: waters XBridge C18, 3.0 x 150mm, 3.5 μm;
mobile phase: adding 5ml of triethanolamine into 1L of methanol aqueous solution with the volume percentage of 42 percent;
detection wavelength: 228 nm;
column temperature: 35 ℃;
sample introduction amount: 10 mu l of the mixture;
flow rate: 1.0 ml/min;
operating time: and (3) 30 min.
1.2 sample preparation
Diluting liquid: 42 percent methanol water solution by volume, and 5ml of triethanolamine is added into 1L of methanol water solution.
Test solution: taking 10mg of the raltitrexed bulk drug in example 1, adding the diluent to dissolve and dilute the raltitrexed bulk drug into a 20ml measuring flask, and shaking up the solution to be used as a test solution.
Control solution: precisely measuring 1ml of the test solution, placing the test solution in a 200ml measuring flask, diluting the test solution to a scale with the diluent solution, and shaking up to obtain a control solution.
Raltitrexed enantiomer stock solutions: taking about 25mg of raltitrexed enantiomer, precisely weighing, placing in a 500ml measuring flask, adding a diluent to dissolve and dilute to a scale, shaking up, and taking as an optical isomer stock solution.
Enantiomeric positioning solution: precisely measuring 5ml of optical isomer stock solution, placing the optical isomer stock solution into a 50ml measuring flask, diluting the optical isomer stock solution to a scale with a diluent, and shaking up to obtain an enantiomer positioning solution.
Resolution solution: taking about 10mg of raltitrexed reference substance, precisely weighing, placing in a 20ml measuring flask, precisely weighing 1ml of raltitrexed enantiomer stock solution, adding diluent to dilute to scale, shaking up, and using as a resolution solution.
1.3 determination of
Injecting 10 μ l of the separation degree solution into a high performance liquid chromatograph, and recording a chromatogram, wherein the separation degree should be more than 1.5.
And respectively injecting 10 mu l of the test solution and 10 mu l of the control solution into a high performance liquid chromatograph, and recording the chromatogram. The enantiomeric contents were calculated according to the self-control method.
The high performance liquid chromatography result of the resolution solution is shown in figure 4, the high performance liquid chromatography result of the test solution is shown in figure 5, the high performance liquid chromatography result of the isomer solution is shown in figure 6, and the raltitrexed and the enantiomer thereof are almost overlapped under the condition, and meanwhile, the figure shows that the triethanolamine has alkalinity, can form ester with the raltitrexed, and has obvious impurities within about 10.6 min.
The triethanolamine solution in the mobile phase is replaced by 0.8 percent g/ml sulfonated beta-cyclodextrin aqueous solution, other chromatographic conditions are unchanged, the result is shown in figure 7, and the chromatographic peaks of raltitrexed and enantiomers thereof are overlapped and have poor peak shape; the proportion of the organic phase in the mobile phase is reduced, the optimization result is shown in figure 8, and the separation sign of the chromatographic peaks of the raltitrexed and the enantiomer thereof can be seen, but the baseline separation is difficult to achieve.
This proves that the liquid phase method of CN107167535A is not suitable for the detection and separation of raltitrexed and its enantiomer, and meanwhile, the chromatographic parameters such as the composition of chromatographic column and mobile phase in the present invention play a key role in the detection and separation of raltitrexed and its enantiomer.
Although the embodiments disclosed in the present application are described above, the descriptions are only for the convenience of understanding the present application, and are not intended to limit the present application. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the disclosure as defined by the appended claims.
Claims (14)
1. A separation and detection method for enantiomers in raltitrexed comprises the following steps:
1) adopting high performance liquid chromatography, setting the chromatographic conditions as follows:
a chromatographic column: octadecylsilane chemically bonded silica gel column is used as a stationary phase;
detection wavelength: 224nm to 230 nm;
column temperature: c, 3-42 ℃;
sample introduction amount: 1 to 20 mu l;
flow rate: 0.8ml/min-1.2 ml/min;
mobile phase: the composite material consists of a mobile phase A and a mobile phase B, wherein the mobile phase A is an aqueous solution of a cyclodextrin derivative, the cyclodextrin derivative in the aqueous solution of the cyclodextrin derivative is sulfonated-beta-cyclodextrin, and the pH value is adjusted to 3.6-6.0 by phosphoric acid; the mobile phase B is acetonitrile; the volume ratio of the mobile phase A to the mobile phase B is (910-930) to (70-90); and carrying out isocratic elution;
2) preparing a test solution:
taking a proper amount of the raltitrexed raw material medicine or the raltitrexed preparation, adding 0.1mol/L sodium hydroxide solution for dissolving and adding water for diluting to prepare a raltitrexed test sample solution;
3) measuring by high performance liquid chromatography under the chromatographic condition of step 1), precisely measuring the sample solution, injecting into a liquid chromatograph, and recording the chromatogram.
2. The separation detection method according to claim 1, wherein the concentration of the cyclodextrin derivative in the aqueous solution of the cyclodextrin derivative is 0.0005 to 0.02 g/ml.
3. The separation detection method according to claim 1, wherein the mobile phase A is an aqueous solution of sulfonated- β -cyclodextrin, and the concentration of sulfonated- β -cyclodextrin in the aqueous solution of sulfonated- β -cyclodextrin is 0.0005 to 0.02 g/ml.
4. The separation detection method according to claim 3, wherein the concentration of the sulfonated- β -cyclodextrin in the aqueous solution of the sulfonated- β -cyclodextrin is 0.006 to 0.012 g/ml.
5. The separation detection method according to claim 4, wherein the concentration of the sulfonated- β -cyclodextrin in the aqueous solution of the sulfonated- β -cyclodextrin is 0.008 g/ml.
6. The separation detection method according to claim 4, wherein the concentration of the sulfonated- β -cyclodextrin in the aqueous solution of the sulfonated- β -cyclodextrin is 0.009 g/ml.
7. The separation detection method according to claim 4, wherein the concentration of the sulfonated- β -cyclodextrin in the aqueous solution of the sulfonated- β -cyclodextrin is 0.01 g/ml.
8. The separation detection method according to claim 1, wherein the mobile phase a is adjusted to pH 5.0 with phosphoric acid.
9. The separation detection method according to claim 1, wherein the volume ratio of the mobile phase a to the mobile phase B is 920: 80.
10. The separation detection method according to claim 1, wherein the detection wavelength is 226 nm;
optionally, the column temperature is 40 ℃;
optionally, the sample size is 10 μ l;
optionally, the flow rate is 1.0 ml/min.
11. The separation detection method according to any one of claims 1 to 10, wherein the concentration of the test solution is 0.5mg/ml to 2 mg/ml.
12. The separation detection method according to claim 11, wherein the concentration of the sample solution is 0.5 mg/ml.
13. The separation detection method according to any one of claims 1 to 10, wherein the step 2) further comprises preparing a control solution: precisely measuring 1ml of the test solution, placing the test solution in a 200ml measuring flask, diluting the test solution to a scale with water, and shaking up to obtain a control solution.
14. The separation detection method according to any one of claims 1 to 10, further comprising preparing a resolution solution in step 2): taking about 10mg of raltitrexed reference substance, precisely weighing, placing in a 20ml measuring flask, precisely weighing 1ml of raltitrexed enantiomer stock solution, adding 2ml of 0.1mol/L sodium hydroxide solution for dissolving, adding water for diluting to a scale, shaking up, and taking as a resolution solution; wherein, the preparation of the raltitrexed enantiomer stock solution comprises the following steps: about 25mg of raltitrexed enantiomer is taken, precisely weighed, placed in a 500ml measuring flask, dissolved by adding 2ml of 0.1mol/L sodium hydroxide solution, diluted to the scale by adding water and shaken up to be used as optical isomer stock solution.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101221147A (en) * | 2007-09-25 | 2008-07-16 | 扬子江药业集团有限公司 | Method for detecting Raltitrexed enantiomer by capillary tube electrophoresis |
| CN107167535A (en) * | 2017-07-20 | 2017-09-15 | 南京普氟生物检测技术有限公司 | A kind of method that reversed phase liquid chromatography detects Raltitrexed enantiomter |
| JP2017528684A (en) * | 2014-05-23 | 2017-09-28 | インスティチュート ナショナル デ ラ サンテ エ デ ラ ルシェルシュ メディカル （インセルム） | How to determine if a patient will get a response after radiation therapy |
| CN109283279A (en) * | 2017-07-21 | 2019-01-29 | 南京正大天晴制药有限公司 | Pass through high efficiency liquid chromatography for separating and determining Raltitrexed and its method of enantiomter |
| CN110749692A (en) * | 2019-10-15 | 2020-02-04 | 扬子江药业集团有限公司 | Separation and detection method of L-glutamic acid diethyl ester hydrochloride and optical isomer thereof |
-
2020
- 2020-04-30 CN CN202010362885.5A patent/CN111678998B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101221147A (en) * | 2007-09-25 | 2008-07-16 | 扬子江药业集团有限公司 | Method for detecting Raltitrexed enantiomer by capillary tube electrophoresis |
| JP2017528684A (en) * | 2014-05-23 | 2017-09-28 | インスティチュート ナショナル デ ラ サンテ エ デ ラ ルシェルシュ メディカル （インセルム） | How to determine if a patient will get a response after radiation therapy |
| CN107167535A (en) * | 2017-07-20 | 2017-09-15 | 南京普氟生物检测技术有限公司 | A kind of method that reversed phase liquid chromatography detects Raltitrexed enantiomter |
| CN109283279A (en) * | 2017-07-21 | 2019-01-29 | 南京正大天晴制药有限公司 | Pass through high efficiency liquid chromatography for separating and determining Raltitrexed and its method of enantiomter |
| CN110749692A (en) * | 2019-10-15 | 2020-02-04 | 扬子江药业集团有限公司 | Separation and detection method of L-glutamic acid diethyl ester hydrochloride and optical isomer thereof |
Non-Patent Citations (3)
| Title |
|---|
| Chiral separation of raltitrexed by cyclodextrin-modified micellar electrokinetic chromatography;Y. Liu et al.;《Analytical and Bioanalytical Chemistry》;20081019;第393卷(第1期);321-326 * |
| 磺丁基醚-β-环糊精流动相添加法拆分尼莫地平对映体;刘尧 等;《世界最新医学信息文摘》;20190830;第19卷(第70期);17-18 * |
| 高效液相色谱手性流动相添加法拆分阿卓乳酸对映体;张虎 等;《色谱》;20140608;第32卷(第6期);612-615 * |
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