阅读说明：本技术 一种(s)-3-羟基四氢呋喃对映异构体的高效液相拆分方法 (High performance liquid phase resolution method of (S) -3-hydroxytetrahydrofuran enantiomer ) 是由 张洪飞 郭赛 王洁 黄治超 于 2020-11-06 设计创作，主要内容包括：本发明公开了一种(S)-3-羟基四氢呋喃对映异构体的高效液相拆分方法,该方法包括用衍生剂将(S)-3-羟基四氢呋喃样品衍生化,再用HPLC法进行测定,其中HPLC法中的流动相为体积比为45：55的水(A)和乙腈(B)。本发明提供了一种快捷、简便的(S)-3-羟基四氢呋喃对映异构体的衍生以及拆分方法,具有测定方法操作简单、检测快速且准确、成本低、耗时短等优势,对(S)-3-羟基四氢呋喃对映异构体的检测,可用于监测以(S)-3-羟基四氢呋喃为原料的恩格列净等原料药及制剂的质量。(The invention discloses a high performance liquid phase resolution method of (S) -3-hydroxytetrahydrofuran enantiomer, which comprises the steps of derivatizing an (S) -3-hydroxytetrahydrofuran sample by using a derivatizing agent and then measuring by using an HPLC method, wherein the mobile phase in the HPLC method is a compound with a volume ratio of 45: 55 of water (A) and acetonitrile (B). The invention provides a rapid, simple and convenient method for deriving and splitting an (S) -3-hydroxytetrahydrofuran enantiomer, which has the advantages of simple operation of a determination method, rapid and accurate detection, low cost, short time consumption and the like, and can be used for monitoring the quality of raw material medicines and preparations such as exellene taking the (S) -3-hydroxytetrahydrofuran as a raw material by detecting the (S) -3-hydroxytetrahydrofuran enantiomer.)

1. A high performance liquid phase resolution method of an enantiomer of (S) -3-hydroxytetrahydrofuran is characterized by comprising the steps of derivatizing a (S) -3-hydroxytetrahydrofuran sample by using a derivatizing agent and measuring by using an HPLC method, wherein the HPLC method comprises the following steps of: 55 of water (A) and acetonitrile (B).

2. The high performance liquid phase resolution method of claim 1, wherein the detection conditions of the HPLC method comprise: the chromatographic column is a xylonite chemical bonding type chiral chromatographic column, the column temperature is 30 ℃, the flow rate is 1.0ml/min, and the detection wavelength is 254 nm.

3. The high performance liquid resolution method of claim 2, wherein the type of the xylonite chemically bonded chiral chromatographic column is CHIRALPAK IA-3, and the specification of the chromatographic column is 4.6 x 250mm x 3 μm.

4. The high performance liquid resolution method of claim 1, wherein the derivatizing agent is 4-nitrobenzenesulfonyl chloride.

5. The high performance liquid resolution process of claim 4 wherein said derivatization is carried out in the presence of a catalyst and pyridine.

6. The high performance liquid resolution process of claim 5 wherein the catalyst is 4-dimethylaminopyridine.

7. The high performance liquid phase resolution method of claim 4, wherein the derivatization time is 30 min.

8. The high performance liquid resolution method according to any one of claims 1 to 7, which comprises the following steps:

(1) preparing a solution: respectively preparing a blank solution, a test solution, a reference solution and a system applicability solution; wherein the test solution comprises a (S) -3-hydroxytetrahydrofuran sample, pyridine and acetonitrile; the reference solution comprises a (R) -3-hydroxytetrahydrofuran sample, pyridine and acetonitrile; the system applicability solution comprises a (S) -3-hydroxytetrahydrofuran sample, a (R) -3-hydroxytetrahydrofuran sample, pyridine and acetonitrile;

(2) and (3) HPLC determination: the prepared solutions were injected into a chromatograph, and chromatograms were recorded.

9. The method for high performance liquid resolution of claim 8, wherein the sample solution is prepared by the following steps: adding 0.5ml of pyridine into a 100g of (S) -3-hydroxytetrahydrofuran sample, then respectively adding 6mg of catalyst and 325mg of derivatization agent, supplementing pyridine to 5ml, derivatizing at room temperature for 30min, placing 1ml of the derivatized sample into a 10ml measuring flask, diluting acetonitrile to scale, filtering, and taking the subsequent filtrate as a sample solution.

10. The high performance liquid phase resolution method of claim 8, wherein the control solution is prepared by the following steps: adding 0.5ml of pyridine into a 100mg (R) -3-hydroxytetrahydrofuran sample, then respectively adding 6mg of catalyst and 325mg of derivatization agent, supplementing pyridine to 5ml, derivatizing at room temperature for 30min, placing 1ml of the derivatized sample into a 10ml measuring flask, diluting acetonitrile to scale, filtering, and taking the subsequent filtrate as a sample solution;

the preparation method of the system applicability solution comprises the following steps: weighing a proper amount of (S) -3-hydroxytetrahydrofuran and a proper amount of (R) -3-hydroxytetrahydrofuran respectively, adding 0.5ml of pyridine, then adding 6mg of catalyst and 325mg of derivatization agent respectively, adding pyridine to 5ml, derivatizing for 30min at room temperature, placing a proper amount in a 10ml measuring flask after derivatizing is finished, diluting acetonitrile to the scale, filtering, and taking the subsequent filtrate as a sample solution.

Technical Field

The invention relates to the field of drug detection and analysis, and more particularly relates to a high-performance liquid phase resolution method of an (S) -3-hydroxytetrahydrofuran enantiomer.

Background

(S) -3-hydroxytetrahydrofuran of the formula C₄H₈O₂The compound is an important pharmaceutical and chemical intermediate, can be used as a raw material of raw material medicaments such as engeletin, afatinib, amprenavir and the like, and has wide application in the synthesis of medicaments such as anti-cancer medicaments, hypoglycemic medicaments, AIDS medicaments and the like.

The existing (S) -3-hydroxytetrahydrofuran often contains enantiomer impurity (R) -3-hydroxytetrahydrofuran, which inevitably causes the quality reduction of raw material medicines such as exendin and the like prepared by taking (S) -3-hydroxytetrahydrofuran as raw materials, and anti-cancer medicines and other medicines, and even causes the problem of medication safety, therefore, the control of the quality of (S) -3-hydroxytetrahydrofuran has important significance.

However, (S) -3-hydroxytetrahydrofuran and its enantiomeric impurity (R) -3-hydroxytetrahydrofuran are very difficult to separate, and there are few documents on quality control of (S) -3-hydroxytetrahydrofuran at present. For example, patent CN110618208A discloses a method for detecting the content of (S) -3-hydroxytetrahydrofuran enantiomer, which comprises derivatizing a sample of (S) -3-hydroxytetrahydrofuran with N, N-dimethylaminopyridine as a catalyst, triethylamine as an acid-binding agent, and acetic anhydride as a derivatizing agent, and performing assay analysis by gas chromatography, and discloses that (S) - (+) -3-hydroxytetrahydrofuran is only absorbed at a terminal wavelength of 193nm by ultraviolet scanning, and therefore, the enantiomer impurity (R) - (+) -3-hydroxytetrahydrofuran in (S) - (+) -3-hydroxytetrahydrofuran cannot be detected by high performance liquid chromatography, and thus, the gas chromatography detection method is adopted in the patent, compared with high performance liquid chromatography, the precision determination standard of the gas chromatography is usually that RSD is less than 10%, and the experimental error is relatively large; the temperature needs to be raised during operation, the injection port and the detector are generally not lower than 200 ℃, and the column temperature box can be as high as nearly 200 ℃ sometimes; meanwhile, the gas phase equipped with the FID also requires a hydrogen source to sustain the detector flame, which is dangerous for both the cylinder and the generator, and the detection takes a long time and has limited practical value, which limits the application of this method to a great extent.

Therefore, it is urgent to find a method for separating the enantiomeric impurities of (S) -3-hydroxytetrahydrofuran, which is easy to operate, and which can be performed rapidly, efficiently and simply.

Disclosure of Invention

The invention aims to overcome the defects and shortcomings in the prior art and provide a high-efficiency liquid phase resolution method of an enantiomer of (S) -3-hydroxytetrahydrofuran.

The above purpose of the invention is realized by the following technical scheme:

a high performance liquid phase resolution method of an enantiomer of (S) -3-hydroxytetrahydrofuran, which comprises derivatizing a sample of (S) -3-hydroxytetrahydrofuran with a derivatizing agent and then measuring by an HPLC method, wherein the HPLC method comprises the following steps of: 55 of water (A) and acetonitrile (B).

The (S) -3-hydroxytetrahydrofuran sample according to the invention is a commercial product which contains a small amount of its enantiomer (R) -3-hydroxytetrahydrofuran.

Preferably, the detection conditions of the HPLC method include: the chromatographic column is a xylonite chemical bonding type chiral chromatographic column, the column temperature is 30 ℃, the flow rate is 1.0ml/min, and the detection wavelength is 254 nm.

Preferably, the type of the xylonite chemically bonded chiral chromatographic column is CHIRALPAK IA-3, and the specification of the chromatographic column is 4.6 mm by 250mm by 3 μm.

Preferably, the derivatizing agent is 4-nitrobenzenesulfonyl chloride.

Preferably, the derivatization is carried out in the presence of a catalyst and pyridine.

Preferably, the catalyst is 4-dimethylaminopyridine.

Preferably, the derivatization time is 30 min.

Preferably, the high performance liquid phase resolution method specifically comprises the following steps:

(1) preparing a solution: respectively preparing a blank solution, a test solution, a reference solution and a system applicability solution; wherein the test solution comprises a (S) -3-hydroxytetrahydrofuran sample, pyridine and acetonitrile;

(2) and (3) HPLC determination: the prepared solutions were injected into a chromatograph, and chromatograms were recorded.

Preferably, the preparation method of the test solution comprises the following steps: adding 0.5ml of pyridine into a 100g of (S) -3-hydroxytetrahydrofuran sample, then respectively adding 6mg of catalyst and 325mg of derivatization agent, supplementing pyridine to 5ml, derivatizing at room temperature for 30min, placing 1ml of the derivatized sample into a 10ml measuring flask, diluting acetonitrile to scale, filtering, and taking the subsequent filtrate as a sample solution.

Preferably, the preparation method of the control solution comprises the following steps: adding 0.5ml of pyridine into a 100mg (R) -3-hydroxytetrahydrofuran sample, then respectively adding 6mg of catalyst and 325mg of derivatization agent, supplementing pyridine to 5ml, derivatizing at room temperature for 30min, placing 1ml of the derivatized sample into a 10ml measuring flask, diluting acetonitrile to scale, filtering, and taking the subsequent filtrate as a sample solution;

the preparation method of the system applicability solution comprises the following steps: weighing a proper amount of (S) -3-hydroxytetrahydrofuran and a proper amount of (R) -3-hydroxytetrahydrofuran respectively, adding 0.5ml of pyridine, then adding 6mg of catalyst and 325mg of derivatization agent respectively, adding pyridine to 5ml, derivatizing for 30min at room temperature, placing a proper amount in a 10ml measuring flask after derivatizing is finished, diluting acetonitrile to the scale, filtering, and taking the subsequent filtrate as a sample solution.

Compared with the prior art, the invention has the following beneficial effects:

the invention provides a method for separating (S) -3-hydroxytetrahydrofuran enantiomer impurities by an HPLC method for the first time, the chromatographic peak separation degree of the (S) -3-hydroxytetrahydrofuran enantiomer by the method is as high as 3.0 and far greater than 1.5, and the method also has high specificity and system applicability.

The method utilizes convenient and quick high performance liquid chromatography, has the advantages of simple operation of the determination method, quick and accurate detection, low cost, short time consumption and the like, and can be used for detecting the enantiomer of the (S) -3-hydroxytetrahydrofuran and monitoring the quality of raw material medicines and preparations such as the exellene taking the (S) -3-hydroxytetrahydrofuran as the raw material.

Drawings

FIG. 1 is a graph showing the results of HPLC detection of the white solution in example 1.

FIG. 2 is a diagram showing the results of HPLC analysis of a solution suitable for use in the system of example 1.

FIG. 3 is a graph showing the results of HPLC analysis of the sample solution in example 1.

FIG. 4 is a graph showing the results of HPLC analysis of the control solution in example 1.

Detailed Description

In order to more clearly and completely describe the technical scheme of the invention, the invention is further described in detail by the specific embodiments, and it should be understood that the specific embodiments described herein are only used for explaining the invention, and are not used for limiting the invention, and various changes can be made within the scope defined by the claims of the invention.

The (S) -3-hydroxytetrahydrofuran samples used according to the invention are all commercially available products which contain small amounts of their enantiomer (R) -3-hydroxytetrahydrofuran.

Example 1

A high performance liquid phase resolution method of (S) -3-hydroxytetrahydrofuran enantiomer comprises the following steps:

(1) preparing a solution:

test solution: adding 0.5ml of pyridine into a 100g (S) -3-hydroxytetrahydrofuran sample, respectively adding 6mg of catalyst 4-dimethylaminopyridine and 325mg of derivatization agent 4-nitrobenzenesulfonyl chloride, supplementing pyridine to 5ml, derivatizing at room temperature for 30min, placing 1ml of the derivatized sample in a 10ml measuring flask, diluting acetonitrile to a scale, filtering, and taking the subsequent filtrate as a sample solution;

blank solution: taking 0.5ml of pyridine, adding about 6mg of catalyst 4-dimethylaminopyridine and 325mg of derivatization agent 4-nitrobenzenesulfonyl chloride, supplementing pyridine to 5ml, shaking up, derivatizing at room temperature for 30min, taking 1ml after derivatization, placing in a 10ml measuring flask, diluting acetonitrile to scale, shaking up, filtering, and taking the subsequent filtrate as a blank solution;

control solution: adding 0.5ml of pyridine into a 100mg (R) -3-hydroxytetrahydrofuran sample, then respectively adding 6mg of catalyst 4-dimethylaminopyridine and 325mg of derivatization agent 4-nitrobenzenesulfonyl chloride, supplementing pyridine to 5ml, derivatizing for 30min at room temperature, taking 1ml after derivatization, placing the 1ml into a 10ml measuring flask, diluting acetonitrile to a scale, filtering, and taking the subsequent filtrate as a test sample solution;

preparing a system applicability solution: weighing a proper amount of (S) -3-hydroxytetrahydrofuran and a proper amount of (R) -3-hydroxytetrahydrofuran respectively, adding 0.5ml of pyridine, then adding 6mg of catalyst 4-dimethylaminopyridine and 325mg of derivatization agent 4-nitrobenzenesulfonyl chloride respectively, supplementing pyridine to 5ml, derivatizing for 30min at room temperature, placing a proper amount in a 10ml measuring flask after derivatizing is finished, diluting acetonitrile to scale, filtering, and taking a subsequent filtrate as a test solution;

(2) and (4) HPLC detection:

respectively injecting the test solution, the blank solution, the reference solution and the system applicability solution into a liquid chromatograph, and recording a chromatogram, wherein the chromatogram conditions are as follows:

chromatographic column	CHIRALPAKIA-3，4.6*250mm*3μm
		Column temperature	30℃
Flow rate of flow	1.0ml/min
		Detection wavelength	254nm
Mobile phase	Water (A): acetonitrile (B) ═ 45: 55

Detection results and analysis:

the HPLC detection results of the blank solution, the system applicability solution, the sample solution and the reference solution are respectively shown in the figures 1-4, and it can be seen from the figures that the blank solvent does not interfere with the detection of (S) -3-hydroxytetrahydrofuran and the enantiomer thereof (R) -3-hydroxytetrahydrofuran, and the method has good specificity.

In the attached drawing 2, the separation degree of (R) -3-hydroxytetrahydrofuran and (S) -3-hydroxytetrahydrofuran are respectively shown from left to right, and as can be seen from the drawing, the separation degree is more than 3.0 and is far more than 1.5, and baseline separation can be achieved, so that the method disclosed by the invention can well separate (R) -3-hydroxytetrahydrofuran from (S) -3-hydroxytetrahydrofuran, and can be completely suitable for separating enantiomer impurities of (S) -3-hydroxytetrahydrofuran.

The present invention has been described in detail with reference to the above examples using specific embodiments and experiments, but it will be apparent to those skilled in the art that modifications or improvements can be made thereto without departing from the spirit of the present invention. Accordingly, such modifications and improvements do not depart from the spirit of the invention and are intended to be included within the scope of the invention.