Method for detecting residual DIPEA in sulfonate hydrochloride and application thereof

Abstract

The invention provides a detection method of residual DIPEA in sulfonate hydrochloride and application thereof, the detection method comprises the following steps: (1) mixing a to-be-detected sample with a first diluent to obtain a test solution; (2) sequentially mixing a reference substance with a first diluent and a second diluent to obtain a reference substance solution; and (3) respectively carrying out gas chromatography detection on the test solution and the reference solution, and calculating the content of residual DIPEA in the to-be-detected sample according to the detection result. The detection method provided by the invention can effectively detect the content of the alkaline residual solvent DIPEA to be detected in the acidic substance sulfonate hydrochloride by utilizing the salting-out effect, and has the advantages of high specificity, high sensitivity, high precision, high accuracy, wide linear range and good solution stability.

Description

Technical Field

[0001] The invention belongs to the technical field of analytical chemistry, and particularly relates to a detection method for residual DIPEA in sulfonate hydrochloride and application thereof. Background Art

[0002] With the continuous development of synthesis technology and high-throughput screening of drugs, more and more salt compounds have entered the stage of new drugs or lead compounds. Drugs can improve the physical and chemical properties of drug molecules through salt formation. This method is the simplest and most effective way to improve drugability, directly affecting the solubility, stability, purity and bioavailability of raw materials. However, drug salt formation does not necessarily affect the safety and effectiveness of the drug, so salt formation is an important trend in the development of existing raw materials.

[0003] Residual solvents in APIs refer to organic solvents that are used in the production of APIs or excipients, as well as in the preparation of preparations, and that are not completely removed during the process. According to the toxicity of organic solvents and their harm to the environment, ICH (International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use) divides organic solvents into four categories: avoidance of use, restricted use, low toxicity, and insufficient toxicity. Residual solvents need to be studied because they can affect the safety of products.

[0004] As salt compounds are increasingly used in drug development, quality research on salt compounds has gradually become an important part of analysis, and the detection of residual solvents has also become one of the difficulties. For salt-forming compounds, alkaline residual solvents in acidic salt compounds or acidic residual solvents in alkaline compounds, it is very common to detect whether the residual solvent is completely freed. The development of each salt compound solvent residual method requires multiple attempts to achieve the recovery rate requirement, which is time-consuming and labor-intensive, and the reproducibility and durability are also poor. Therefore, the development of a new method for detecting alkaline residual solvents in acidic salt compounds has become an urgent problem to be solved. Summary of the invention

[0005] In view of the shortcomings of the prior art, the object of the present invention is to provide a detection method for residual DIPEA in sulfonate hydrochloride and its application. The detection method provided by the present invention can effectively detect the content of the alkaline residual solvent DIPEA to be detected in the acidic substance sulfonate hydrochloride by using the salting-out effect, and has the advantages of high specificity, high sensitivity, high precision, high accuracy, wide linear range and good solution stability.

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

[0007] On the one hand, the present invention provides a method for detecting residual DIPEA (diisopropylethylamine) in a sulfonate hydrochloride, the detection method comprising the following steps:

[0008] (1) mixing a sample to be tested with a first diluent to obtain a test solution;

[0009] (2) mixing the reference substance with the first diluent and the second diluent in sequence to obtain a reference substance solution;

[0010] (3) The test solution and the reference solution are respectively subjected to gas chromatography detection, and the content of residual DIPEA in the test sample is calculated based on the detection results.

[0011] There is no order distinction between steps (1) and (2).

[0012] In step (1), the first diluent is an alkaline solution, and in step (2), the second diluent is an acid-base mixed solution.

[0013] The above method utilizes the salting-out effect to release the residual DIPEA in the sample to be tested using an alkaline solution, and uses an acid-base mixed solution to dilute the reference substance, thereby simulating the salt environment in the test solution and improving the accuracy of the detection. Ultimately, the detection method has the advantages of high specificity, high sensitivity, high precision, high accuracy, a wide linear range and good solution stability.

[0014] Preferably, in the alkaline solution, the base comprises any one of triethylamine, diethylamine or 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) or a combination of at least two thereof, preferably 1,8-diazabicyclo[5.4.0]undec-7-ene.

[0015] The above-mentioned specific base can effectively release the residual DIPEA in the sample to be tested, further improving the accuracy of the detection.

[0016] Preferably, in the acid-base mixed solution, the base includes any one of triethylamine, diethylamine or 1,8-diazabicyclo[5.4.0]undec-7-ene or a combination of at least two thereof, and the acid includes any one of hydrochloric acid, sulfuric acid, perchloric acid or trifluoroacetic acid or a combination of at least two thereof.

[0017] Preferably, the solvents in the first diluent and the second diluent are independently selected from any one of NMP (N-methylpyrrolidone), DMF (N,N-dimethylformamide) or DMSO (dimethyl sulfoxide) or a combination of at least two thereof.

[0018] Preferably, the molar ratio of the sulfonate hydrochloride in the sample to be tested to the alkali in the alkali solution is 1:(3-5), such as 1:3, 1:3.5, 1:4, 1:4.5 or 1:5, but is not limited to the above-listed values. Other values ​​not listed within the above numerical range are also applicable.

[0019] Preferably, the molar ratio of acid to base in the acid-base mixed solution is 1:(1-2), such as 1:1, 1:1.2, 1:1.4, 1:1.6, 1:1.8 or 1:2, but is not limited to the above-listed values, and other values ​​not listed within the above numerical range are also applicable.

[0020] Preferably, the chromatographic column for gas chromatography detection in step (3) is CP-Volamine, 60m×320μm.

[0021] Preferably, in the gas chromatography detection in step (3), the column flow rate is 2-5 mL / min, for example 2 mL / min, 2.5 mL / min, 3 mL / min, 3.5 mL / min, 4 mL / min, 4.5 mL / min or 5 mL / min, but is not limited to the above-listed values. Other values ​​not listed within the above numerical range are also applicable.

[0022] Preferably, the temperature rise program of the gas chromatography detection in step (3) is as follows:

[0023] The starting temperature is 90-190°C and maintained for 0.8-1.2 min, then the temperature is increased to 240-260°C at a rate of 10-20°C / min and maintained for 7.5-8.5 min.

[0024] The starting temperature may be 90°C, 100°C, 110°C, 120°C, 130°C, 140°C, 150°C, 160°C, 170°C, 180°C or 190°C, etc., the holding time may be 0.8min, 0.9min, 1min, 1.1min or 1.2min, etc., and the heating rate may be 10°C / min, 11°C / min, 12°C / min, 13°C / min, 14°C / min, 15°C / min, 16°C / min, 17°C / min , 18℃ / min, 19℃ / min or 20℃ / min, etc. The temperature increased to may be 240℃, 245℃, 250℃, 255℃ or 260℃, etc. The holding time may be 7.5min, 7.6min, 7.7min, 7.8min, 7.9min, 8min, 8.1min, 8.2min, 8.3min, 8.4min or 8.5min, etc., but are not limited to the values ​​listed above, and other values ​​not listed within the above numerical range are equally applicable.

[0025] The above-mentioned specific parameter control can effectively improve the separation degree between the sulfonate hydrochloride and DIPEA peaks and improve the detection accuracy.

[0026] On the other hand, the present invention also provides the use of the above detection method in the detection of alkaline residual solvents in acidic substances.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] The invention provides a method for detecting residual DIPEA in sulfonate hydrochloride. By adopting the salting-out effect, an alkaline solution can be used to release the residual DIPEA in a sample to be tested, and an acid-base mixed solution is used to dilute a reference substance, so that the salt environment in a test solution can be simulated, and the accuracy of detection is improved. Finally, the detection method has the advantages of high specificity, high sensitivity, high precision, high accuracy, wide linear range and good solution stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 is a graph showing the test results of the test solution of Example 1;

[0030] Figure 2 It is a graph of the specificity test results;

[0031] Figure 3 is the quantitative limit test result graph;

[0032] Figure 4 is the detection limit test result graph;

[0033] Figure 5 is a test result diagram of Example 2;

[0034] Figure 6 is a test result diagram of Example 3;

[0035] Figure 7 This is a graph of column flow rate study test results;

[0036] Figure 8 This is a graph of the test results of the heating program study. DETAILED DESCRIPTION

[0037] 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.

[0038] In the following examples, the sources of reagents, test drugs and instruments are as follows:

[0039] Gas chromatograph: Model: 8890, Agilent Technologies, Inc.;

[0040] Detector: FID detector, Agilent Technologies Ltd.;

[0041] Chromatographic column: CP-Volamine (60m×320μm), Agilent Technologies;

[0042] Triethylamine (TEA): analytical grade, Aladdin Biochemical Technology Co., Ltd.;

[0043] Diethylamine (DEA): analytical grade, Aladdin Biochemical Technology Co., Ltd.;

[0044] N,N-Diisopropylethylamine (DIPEA): analytical grade, Aladdin Biochemical Technology Co., Ltd.;

[0045] 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), analytical grade, Aladdin Biochemical Technology Co., Ltd.;

[0046] Dimethyl sulfoxide, chromatographic grade, Honeywell Ltd.;

[0047] Concentrated hydrochloric acid, analytical grade, Aladdin Biochemical Technology Co., Ltd.;

[0048] Example 1

[0049] This example provides a method for detecting residual DIPEA in sulfonate hydrochloride, and the specific steps are as follows:

[0050] The sample to be tested was Cerdulatinib hydrochloride (CAS: 1369761-01-2, purchased from Nanjing Yaoshi Technology Co., Ltd.).

[0051] Thinner 1

[0052] DMSO:DBU=95:5(v / v)

[0053] 25 mL of DBU was added to 475 mL of DMSO and mixed to obtain diluent 1.

[0054] Thinner 2

[0055] DMSO:DBU:HCl=200:10:3(v / v / v)

[0056] 20 mL of DBU was added to 400 mL of DMSO, and 6 mL of HCl was added and mixed to obtain diluent 2.

[0057] Dilute 200 mg of the sample to be tested with 2.0 mL of diluent 1 to obtain a test solution;

[0058] Accurately weigh 200 mg of DIPEA into a 100 mL volumetric flask containing 20 mL of diluent 1. Dilute to the mark with diluent 1, mix well, and mark as stock solution 1; transfer 1 mL of stock solution 1 to a 20 mL volumetric flask containing 15 mL of diluent 2, dilute to the mark with diluent 2, mix well, and mark as STD1, i.e., the reference solution. Prepare STD2 in parallel.

[0059] The above test solution and control solution were subjected to headspace gas chromatography using a CP-Volamine column, nitrogen as carrier gas, a column flow rate of 4 mL / min, an inlet temperature of 230°C, a detector temperature of 250°C, a split ratio of 30:1, and a program temperature of 100°C for 1 minute; then raised to 250°C at a rate of 18°C / min and held at this temperature for 8 minutes. The test results of the test solution are as follows: Figure 1 shown.

[0060] According to the test results, it can be calculated that there is no DIPEA residue in the sample.

[0061] Example 2

[0062] This example provides a method for detecting residual DIPEA in sulfonate hydrochloride. The specific steps are the same as those in Example 1 except that DBU is replaced with an equal amount of triethylamine.

[0063] Example 3

[0064] This example provides a method for detecting residual DIPEA in sulfonate hydrochloride. The specific steps are the same as those in Example 1 except that DBU is replaced with an equal amount of diethylamine.

[0065] Example 4

[0066] This example provides a method for detecting residual DIPEA in sulfonate hydrochloride. The specific steps are consistent with those in Example 1 except that the column flow rate is 5 mL / min.

[0067] Example 5

[0068] This example provides a method for detecting residual DIPEA in sulfonate hydrochloride. The specific steps are consistent with those in Example 1 except that the column flow rate is 3 mL / min.

[0069] Example 6

[0070] This example provides a method for detecting residual DIPEA in sulfonate hydrochloride. The specific steps are consistent with those in Example 1 except that the temperature rise procedure is as follows.

[0071] The initial temperature of the program heating was 100°C and maintained for 1 minute; then the temperature was raised to 250°C at a rate of 10°C / min and kept constant for 8 minutes.

[0072] Example 7

[0073] This example provides a method for detecting residual DIPEA in sulfonate hydrochloride. The specific steps are consistent with those in Example 1 except that the temperature rise procedure is as follows.

[0074] The initial temperature of the program heating was 180℃, which was maintained for 1 minute; then the temperature was raised to 250℃ at a rate of 18℃ / min and kept constant for 8 minutes.

[0075] Exclusive properties:

[0076] The diluent 1, diluent 2 and reference solution in Example 1 were injected, and the results are shown in the following table and Figure 2 shown.

[0077]

[0078]

[0079] Limit of Quantitation:

[0080] Accurately transfer 1 mL of the reference solution to a 10 mL volumetric flask containing 2 mL of diluent 2. Dilute to scale with diluent 2, mix well, and mark as sensitivity solution (LOQ). Then, perform the test using the method of Example 1. The results are as follows: Figure 3 As shown in the table below, the sensitivity of the solution signal-to-noise ratio is greater than 10.

[0081]

[0082] Linear:

[0083] Accurately weigh 500 mg of DIPEA into a 50 mL volumetric flask containing 20 mL of diluent 1. Dilute to volume with diluent 1, mix well, and mark as stock solution A. Then prepare the linear solution as follows:

[0084]

[0085]

[0086] Then, the detection was performed according to the method of Example 1 to obtain a standard curve, as shown in the following table:

[0087]

[0088] Detection Limit:

[0089] The detection limit solution was prepared according to the following table, and then the detection was performed according to the method of Example 1:

[0090]

[0091] The results are as follows Figure 4 As shown in the table below, the detection limit solution signal-to-noise ratio is greater than 3:

[0092]

[0093] stability:

[0094] The reference solution in Example 1 was subjected to gas chromatography after being placed for 2 h, 5.5 h, and 7.5 h, and the results were as follows:

[0095]

[0096]

[0097] Accuracy:

[0098] Prepare the spiking solution as follows:

[0099]

[0100] Then the method of Example 1 was used for detection, and the results were as follows:

[0101]

[0102]

[0103] In addition, the same test was performed on the method of Example 2-3, and the results were as follows:

[0104] The detection was performed in the same manner as in Example 1 except that DBU was replaced with an equal amount of triethylamine in Example 2. DIPEA in the reference solution could not be released and could not be detected, and could not be applied to the detection of residual DIPEA in the sulfonate hydrochloride. Figure 5 shown.

[0105] The detection was performed in the same manner as in Example 1 except that DBU was replaced with an equal amount of diethylamine in Example 3. DIPEA in the reference solution was not completely free and produced too many impurity peaks. To avoid limiting the applicability of the method, it should not be used to detect residual DIPEA in the sulfonate hydrochloride. The results are shown in FIG. Figure 6 shown.

[0106] Except that the carrier gas flow rate was adjusted to 5 mL / min and 3 mL / min in Examples 4 and 5, the rest was the same as in Example 1. The flow rate had little effect on the DIPEA peak and separation. Considering the optimal and conventional flow rate suitable for the chromatographic column, 4 mL / min was selected. Figure 7 shown.

[0107] The temperature program in Examples 6 and 7 was followed, and the rest was the same as in Example 1. The initial temperature of the program was 100°C, maintained for 1 minute, and then increased to 250°C at a rate of 10°C / min, and maintained at this temperature for 8 minutes. Figure 8 As shown. The separation degree increases with the decrease of the heating rate, but the corresponding running time will increase; when the initial temperature of the program heating is 180°C, maintained for 1 minute, and then raised to 250°C at a rate of 18°C / min and kept at the constant temperature for 8 minutes, the separation degree does not reach the system suitability requirement of greater than 1.5 (1.42). Therefore, under the premise of ensuring the separation degree and considering the principle of resource conservation, the intermediate rate heating program of Example 1 is selected.

[0108] Precision:

[0109] The reference solution in Example 1 was injected six times continuously, and the results were as follows:

[0110]

[0111] Separation:

[0112]

[0113] The applicant declares that the present invention uses the above-mentioned embodiments to illustrate the detection method and application of residual DIPEA in the sulfonate hydrochloride of the present invention, 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.

[0114] 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.

[0115] 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 residual DIPEA in sulfonate hydrochloride, characterized in that: The detection method comprises the following steps: (1) mixing a sample to be tested with a first diluent to obtain a test solution; (2) mixing the reference substance with the first diluent and the second diluent in sequence to obtain a reference substance solution; (3) The test solution and the reference solution are subjected to gas chromatography detection respectively, and the content of residual DIPEA in the test sample is calculated according to the detection results; Steps (1) and (2) are not sequential; In step (1), the first diluent is an alkaline solution, and in step (2), the second diluent is an acid-base mixed solution.

2. The detection method according to claim 1, characterized in that: In the alkaline solution, the base includes any one of triethylamine, diethylamine or 1,8-diazabicyclo[5.4.0]undec-7-ene or a combination of at least two thereof, preferably 1,8-diazabicyclo[5.4.0]undec-7-ene.

3. The detection method according to claim 1 or 2, characterized in that: In the acid-base mixed solution, the base includes any one of triethylamine, diethylamine or 1,8-diazabicyclo[5.4.0]undec-7-ene or a combination of at least two thereof, and the acid includes any one of hydrochloric acid, sulfuric acid, perchloric acid or trifluoroacetic acid or a combination of at least two thereof.

4. The detection method according to any one of claims 1 to 3, characterized in that: The solvents in the first diluent and the second diluent are independently selected from any one of NMP, DMF or DMSO or a combination of at least two thereof.

5. The detection method according to any one of claims 1 to 4, characterized in that: The molar ratio of the sulfonate hydrochloride in the sample to be tested to the alkali in the alkali solution is 1:(3-5).

6. The detection method according to any one of claims 1 to 5, characterized in that: The molar ratio of the acid to the base in the acid-base mixed solution is 1:(1-2).

7. The detection method according to any one of claims 1 to 6, characterized in that: The chromatographic column for gas chromatography detection in step (3) is CP-Volamine, 60m×320μm.

8. The detection method according to any one of claims 1 to 7, characterized in that: In the gas chromatography detection of step (3), the column flow rate is 2-5 mL / min.

9. The detection method according to any one of claims 1 to 8, characterized in that: The temperature rise program of the gas chromatography detection in step (3) is as follows: The starting temperature is 90-190°C and maintained for 0.8-1.2 min, then the temperature is increased to 240-260°C at a rate of 10-20°C / min and maintained for 7.5-8.5 min.

10. Use of the detection method according to any one of claims 1 to 9 in the detection of alkaline residual solvents in acidic substances.

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