Detection method and application of (2R)-1, 4-dioxane-2-methanol and enantiomer thereof

Abstract

The invention provides a detection method and application of (2R)-1, 4-dioxane-2-methanol and enantiomers thereof, and the detection method comprises the following steps: mixing a sample to be detected with a diluent to obtain a test solution, then carrying out gas chromatography detection, analyzing a sample result according to a detection result, and calculating the content, the enantiomer is (2S)-1, 4-dioxane-2-methanol, and the structural formula of the enantiomer is shown in the description. The detection method provided by the invention can effectively detect the (2S)-1, 4-dioxane-2-methanol in the (2R)-1, 4-dioxane-2-methanol product, the separation degree is high, the method is simple and rapid, and accurate quantification can be realized.

Description

Technical Field

[0001] The invention belongs to the field of analytical chemistry, and specifically relates to a detection method and application of (2R)-1,4-dioxane-2-methanol and its enantiomers, and in particular to a detection method and application of (2R)-1,4-dioxane-2-methanol and its enantiomers with high separation. Background Art

[0002] (2R)-1,4-Dioxane-2-methanol is a common pharmaceutical raw material. It has long been used as the main raw material for the synthesis of some highly active drugs. Studies have shown that it has a good inhibitory activity against the replication of Sindbis virus. In recent years, as people have discovered that it has unique biological activity and that drugs designed based on this structure are effective, the market demand for it has also increased. (2R)-1,4-Dioxane-2-methanol has the following chemical structure:

[0003]

[0004] There is one chiral center in the structure of this compound, so there is only one enantiomer, namely (2S)-1,4-dioxane-2-methanol. Quality control is required in the production process of (2R)-1,4-dioxane-2-methanol, but it is not currently included in the United States Pharmacopoeia USP, the European Pharmacopoeia EP and the Chinese Pharmacopoeia ChP, and no relevant literature has been found to report the use of gradient elution to separate and determine (2R)-1,4-dioxane-2-methanol and its enantiomers. In order to better and more accurately control the content of enantiomers in (2R)-1,4-dioxane-2-methanol and minimize the quality risks in the subsequent use process, it is urgent to develop a new simple, fast and accurate analytical method for determining the content of (2R)-1,4-dioxane-2-methanol enantiomers. Summary of the invention

[0005] In view of the deficiencies in the prior art, the object of the present invention is to provide a method for detecting (2R)-1,4-dioxane-2-methanol and its enantiomers and its application, and in particular to provide a method for detecting (2R)-1,4-dioxane-2-methanol and its enantiomers with high separation and its application. The detection method provided by the present invention can effectively detect (2S)-1,4-dioxane-2-methanol in the (2R)-1,4-dioxane-2-methanol product, has high separation, is simple and fast, and can achieve accurate quantification.

[0006] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0007] In one aspect, the present invention provides a method for detecting (2R)-1,4-dioxane-2-methanol and its enantiomers, the detection method comprising the following steps:

[0008] The sample to be tested is mixed with a diluent to obtain a test solution, which is then tested by gas chromatography. The sample results are analyzed based on the test results, and the content is calculated.

[0009] The enantiomer is (2S)-1,4-dioxane-2-methanol.

[0010] The above method can effectively detect (2S)-1,4-dioxane-2-methanol in the (2R)-1,4-dioxane-2-methanol product with high separation degree. The method is simple and rapid and can achieve accurate quantification.

[0011] The structural formula of the (2R)-1,4-dioxane-2-methanol and its enantiomer (2S)-1,4-dioxane-2-methanol is as follows:

[0012]

[0013] Preferably, the diluent includes any one of acetonitrile, methanol or dichloromethane, or a combination of at least two thereof.

[0014] Preferably, the diluent is acetonitrile.

[0015] Preferably, the chromatographic column for gas phase detection comprises Agilent J&W CycloSil-B or AstecCHIRALDEX B-DM.

[0016] Preferably, the chromatographic column for gas phase detection is Agilent J&W CycloSil-B.

[0017] Preferably, the flow rate of the gas phase detection is 0.8-1.2 mL / min, for example 0.8 mL / min, 0.9 mL / min, 1 mL / min, 1.1 mL / min or 1.2 mL / min, but is not limited to the above-listed values, and other values ​​not listed within the above numerical range are also applicable.

[0018] Preferably, the temperature rise program of the gas phase detection is a gradient temperature rise, and the process of the gradient temperature rise is as follows:

[0019] The starting temperature is 75-85°C, maintained for 1.8-2.2 min; then the temperature is increased to 125-135°C at a rate of 3.5-4.5°C / min, maintained for 1.8-2.2 min; finally, the temperature is increased to 190-210°C at a rate of 28-32°C / min, maintained for 5.5-6.5 min.

[0020] The above-mentioned specific diluent, chromatographic column and temperature program can effectively improve the separation degree of detection and improve the detection accuracy.

[0021] Preferably, the concentration of the test solution is 5-15 mg / mL, for example 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 11 mg / mL, 12 mg / mL, 13 mg / mL, 14 mg / mL or 15 mg / mL, but is not limited to the above-listed values, and other values ​​not listed within the above numerical range are also applicable.

[0022] Preferably, in the gas phase inspection, the injection port temperature is 180-220°C, for example, 180°C, 185°C, 190°C, 195°C, 200°C, 205°C, 210°C, 215°C or 220°C, but is not limited to the above-listed values, and other unlisted values ​​within the above numerical range are also applicable.

[0023] On the other hand, the present invention also provides the use of the above detection method in the quality control of (2R)-1,4-dioxane-2-methanol.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] The invention provides a method for detecting (2R)-1,4-dioxane-2-methanol and its enantiomers, which can effectively detect (2S)-1,4-dioxane-2-methanol in a (2R)-1,4-dioxane-2-methanol product, has high separation degree, is simple and rapid, and can achieve accurate quantification. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 is the gas chromatogram of the blank solution in Example 1;

[0027] Figure 2 is a gas chromatogram of the system suitability solution of (2R)-1,4-dioxane-2-methanol and its enantiomers in Example 1;

[0028] Figure 3 is the gas chromatogram of the sensitivity solution in Example 1;

[0029] Figure 4 It is the gas chromatogram of the test solution in Example 1. DETAILED DESCRIPTION

[0030] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0031] Example 1

[0032] This embodiment provides a method for detecting (2R)-1,4-dioxane-2-methanol and its enantiomers, which is as follows:

[0033] 1. Instruments and conditions

[0034] Agilent 8890 gas chromatograph;

[0035] XSR 205DU electronic analytical balance (Mettler Toledo);

[0036] The raw material (2R)-1,4-dioxane-2-methanol was from Nanjing Yaoshi Technology Co., Ltd.;

[0037] (2R)-1,4-dioxane-2-methanol reference substance and its enantiomer reference substance were purchased from Shanghai Bid Pharmaceutical Technology Co., Ltd.;

[0038] Chromatographic column: CycloSil-B (30m×0.25mm×0.25μm) was purchased from Agilent Technologies.

[0039] 2. Experimental Methods and Results

[0040] 1. Chromatographic conditions

[0041] Chromatographic column: Agilent J&W CycloSil-B (30m×0.25mm×0.25μm);

[0042] Flow rate: 1.0 mL / min (constant flow mode);

[0043] Injection volume: 1 μL;

[0044] Split ratio: 40:1;

[0045] Inlet temperature: 200°C;

[0046] FID detector temperature: 200°C;

[0047] Carrier gas: nitrogen.

[0048] Gradient heating method and other parameters are as follows:

[0049] Gradient heating method of gas phase

[0050]

[0051] 2. Solution Preparation

[0052] Diluent: acetonitrile.

[0053] System suitability solution: Take 50 mg of (2R)-1,4-dioxane-2-methanol reference substance and about 15 mg of (2S)-1,4-dioxane-2-methanol reference substance, accurately weigh, place in a 5 mL volumetric flask, add diluent to dilute to the scale, shake well, and obtain ((2R)-1,4-dioxane-2-methanol concentration 10 mg / mL, (2S)-1,4-dioxane-2-methanol concentration 3 mg / mL).

[0054] Enantiomer reference solution: Take 50 mg of (2S)-1,4-dioxane-2-methanol reference, weigh accurately, place in a 5 mL volumetric flask, dilute to scale with diluent, and shake well. Take 1 mL of the above solution and place in a 100 mL volumetric flask, dilute to scale with diluent, and shake well to obtain ((2S)-1,4-dioxane-2-methanol concentration 0.1 mg / mL).

[0055] Sensitivity solution: Accurately measure 1 mL of the enantiomer reference solution and place it in a 20 mL volumetric flask. Add diluent to dilute to the mark and shake well to obtain ((2S)-1,4-dioxane-2-methanol concentration 0.005 mg / mL).

[0056] Test sample solution: Take 50 mg of the test sample, accurately weigh it, place it in a 5 mL volumetric flask, add an appropriate amount of diluent to dissolve it, add diluent to dilute to the scale, shake well, and you have (test sample concentration 10 mg / mL).

[0057] 3. Exclusivity

[0058] The diluent, system suitability solution, test solution and blank solution (acetonitrile only) were injected and tested respectively. The results showed that the retention time (RT) of the peak position of (2R)-1,4-dioxane-2-methanol was about 13.1, the retention time (RT) of its enantiomer was about 13.8, the separation degree reached 4.0, and the blank did not interfere with the determination. The blank solution chromatogram is shown in Figure 1 (The horizontal axis is time, the vertical axis is signal intensity, the same as the following figures), the system suitability chromatogram is shown in Figure 2 .

[0059] 4. Sensitivity

[0060] The sensitivity solution was used for detection. The results showed that under this concentration condition, the signal-to-noise ratio of the chromatographic peak of (2S)-1,4-dioxane-2-methanol was 10.3, with high sensitivity. The chromatogram of the sensitivity solution is shown in Figure 3 .

[0061] 5. Linear

[0062] Weigh the (2S)-1,4-dioxane-2-methanol reference substance accurately, dissolve it in the diluent, dilute and shake it well, prepare a series of concentration solutions, inject them into the gas chromatograph, perform linear regression, and obtain the regression equation of (2S)-1,4-dioxane-2-methanol. The results show that within the concentration range of 0.005-0.3mg / mL (i.e. 5% to 300% of the limit concentration, (2S)-1,4-dioxane-2-methanol limit is 1.0%, and the limit concentration is 0.1mg / mL), the linear relationship is good, R=0.9998.

[0063] 6. Precision

[0064] The above enantiomer reference solution was sampled and tested continuously, and the chromatogram was recorded. The RSD of the peak area of ​​(2S)-1,4-dioxane-2-methanol was less than 5.0% (n=5), indicating that the precision of this method was good.

[0065]

[0066] 7. Accuracy

[0067] Prepare the (2S)-1,4-dioxane-2-methanol spiked accuracy solutions with a limit concentration of 50%, 100%, and 150% (limit concentration of 0.1 mg / mL (2S)-1,4-dioxane-2-methanol) and calculate the recovery of (2S)-1,4-dioxane-2-methanol. The results show that the recovery of (2S)-1,4-dioxane-2-methanol spiked accuracy is between 80% and 120%, indicating that the accuracy of this method is good.

[0068]

[0069] 8. Stability

[0070] The above 100% limit concentration spiked accuracy solution and enantiomer reference solution were placed at room temperature, sampled and measured at 12 hours, and the chromatograms were recorded. The ratio of the peak area of ​​(2S)-1,4-dioxane-2-methanol in the 100% limit concentration spiked accuracy solution and enantiomer reference solution placed at room temperature for 12 hours to the peak area of ​​(2S)-1,4-dioxane-2-methanol in the 100% limit concentration spiked accuracy solution and enantiomer reference solution at 0 hours was between 80% and 120%, indicating that (2S)-1,4-dioxane-2-methanol was stable after being placed at room temperature for 12 hours.

[0071] 9. Determination of (2S)-1,4-dioxane-2-methanol content in raw materials

[0072] Prepare (2S)-1,4-dioxane-2-methanol reference solution and (2R)-1,4-dioxane-2-methanol test solution according to the above method, inject and measure respectively, and record the chromatogram. If there is a peak in the test solution with the same retention time as the reference solution, integrate this peak as (2S)-1,4-dioxane-2-methanol, and calculate the content of (2S)-1,4-dioxane-2-methanol by peak area according to the external standard method. See the chromatogram of the test solution for details. Figure 4 The content of (2S)-1,4-dioxane-2-methanol in the test samples was 0.39%.

[0073] Example 2

[0074] This embodiment provides a method for detecting (2R)-1,4-dioxane-2-methanol and its enantiomers. The specific method is consistent with that in Example 1 except that the chromatographic column is replaced with Supelco GAMMA-DEX 225.

[0075] Example 3

[0076] This embodiment provides a method for detecting (2R)-1,4-dioxane-2-methanol and its enantiomers. The specific method is consistent with that in Embodiment 1 except that the diluent is replaced by methanol.

[0077] Example 4

[0078] This embodiment provides a method for detecting (2R)-1,4-dioxane-2-methanol and its enantiomers. The specific method is consistent with that in Embodiment 1 except that the diluent is replaced by dichloromethane.

[0079] Example 5

[0080] This embodiment provides a method for detecting (2R)-1,4-dioxane-2-methanol and its enantiomers. The specific method is consistent with that of Example 1 except that the following heating method is used.

[0081]

[0082] Example 6

[0083] This example provides a method for detecting (2R)-1,4-dioxane-2-methanol and its enantiomers. The specific method is consistent with Example 1 except that the flow rate is changed to 2.0 mL / min.

[0084] Effect comparison:

[0085] The detection separations of the methods provided in Examples 1-6 are summarized, and the results are as follows:

[0086] Group Example 1 Example 2 Example 3 Separation 4.0 0.3 3.7 Group Example 4 Example 5 Example 6 Separation 3.8 2.8 3.7

[0087] From the above data, it can be found that the method provided by the present invention can effectively analyze (2R)-1,4-dioxane-2-methanol and its enantiomers; and by selecting a specific chromatographic column, diluent, heating program and flow rate, the detection separation can be effectively improved and the detection accuracy can be improved.

[0088] The applicant declares that the present invention uses the above-mentioned embodiments to illustrate the detection method and application of (2R)-1,4-dioxane-2-methanol and its enantiomers, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0089] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0090] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A method for detecting (2R)-1,4-dioxane-2-methanol and its enantiomers, characterized in that: The detection method comprises the following steps: The sample to be tested is mixed with a diluent to obtain a test solution, which is then tested by gas chromatography, and the sample results are analyzed based on the test results, and the content is calculated; The enantiomer is (2S)-1,4-dioxane-2-methanol.

2. The detection method according to claim 1, characterized in that: The diluent includes any one of acetonitrile, methanol or dichloromethane or a combination of at least two thereof.

3. The detection method according to claim 2, characterized in that: The diluent is acetonitrile.

4. The detection method according to any one of claims 1 to 3, characterized in that: The chromatographic column for gas phase detection includes Agilent J&W CycloSil-B or Supelco GAMMA-DEX 225.

5. The detection method according to claim 4, characterized in that: The chromatographic column for gas phase detection is Agilent J&W CycloSil-B.

6. The detection method according to any one of claims 1 to 5, characterized in that: The flow rate of the gas phase detection is 0.8-1.2 mL / min.

7. The detection method according to any one of claims 1 to 6, characterized in that: The temperature rise procedure of the gas phase detection is a gradient temperature rise, and the process of the gradient temperature rise is as follows: The starting temperature is 75-85°C, maintained for 1.8-2.2 min; then the temperature is increased to 125-135°C at a rate of 3.5-4.5°C / min, maintained for 1.8-2.2 min; finally, the temperature is increased to 190-210°C at a rate of 28-32°C / min, maintained for 5.5-6.5 min.

8. The detection method according to any one of claims 1 to 7, characterized in that: The concentration of the test solution is 5-15 mg / mL.

9. The detection method according to any one of claims 1 to 8, characterized in that: In the gas phase test, the injection port temperature is 180-220°C.

10. Use of the detection method according to any one of claims 1 to 9 in the quality control of (2R)-1,4-dioxane-2-methanol.

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