A method for separating and detecting 1,3-propane sultone

Abstract

The present invention relates to a method for separating and detecting 1,3-propane sultone, and belongs to the field of analytical chemistry. The method comprises: using a chromatographic column with octadecylsilane bonded phase filler, wherein the mobile phase is divided into phase A and phase B, wherein phase A is an aqueous solution of ammonia water or an ammonium salt, and phase B is acetonitrile. The method of the present invention has high sensitivity, good specificity, and reliable accuracy.

Description

Technical Field

[0001] The invention relates to the field of analytical chemistry, and in particular to a method for separating and detecting 1,3-propane sultone. Background Art

[0002] 1,3-Propane sultone, CAS: 1120-71-4, has a chemical structure as shown in Formula I below:

[0003]

[0004] According to the preliminary compilation and reference of the list of carcinogens published by the World Health Organization's International Agency for Research on Cancer on October 27, 2017, 1,3-propane sultone is on the list of Class 2A carcinogens. Therefore, in the drug synthesis process, the content of 1,3-propane sultone needs to be strictly controlled, otherwise it will have a significant impact on drug safety.

[0005] At present, the United States Pharmacopoeia (USP), the European Pharmacopoeia (EP), the Japanese Pharmacopoeia (JP) and the Chinese Pharmacopoeia (Ch.P.) do not include the corresponding separation and detection methods, and there is no existing technology that discloses its detection method. In order to better and more accurately control the impurities in the product and ensure the quality of the drug, it is necessary to study the analysis method suitable for 1,3-propane sultone. Summary of the invention

[0006] The purpose of the present invention is to provide a method for separating and detecting 1,3-propane sultone. The method of the present invention can effectively separate and determine the content of 1,3-propane sultone, the method has high sensitivity, good specificity, reliable accuracy, and can be used for quality control of 1,3-propane sultone in drugs.

[0007] A method for separating and detecting 1,3-propane sultone comprises: using a chromatographic column with octadecylsilane bonded phase filler, wherein the mobile phase is divided into phase A and phase B, wherein phase A is ammonia water or an aqueous solution of ammonium salt, and phase B is acetonitrile.

[0008] In some embodiments, the filler material of the chromatography column is ethylene bridged hybrid particles (BEH).

[0009] The chromatographic column is selected from Waters Xbridge C18 column or Waters XBridge TM In some embodiments, the chromatographic column is a Waters Xbridge C18 column purchased from Waters.

[0010] The ammonium salt is selected from one of ammonium formate and ammonium acetate. In some embodiments, the phase A is selected from ammonia water, which is beneficial to the separation and detection.

[0011] In some embodiments, the volume ratio of the ammonia water or ammonium salt to water is 5:10000 to 2:1000 by volume. In some embodiments, the volume ratio of the ammonia water to water is 1:1000.

[0012] In some embodiments, the volume ratio of phase A to phase B is 95:5 to 5:95. In some embodiments, the volume ratio of phase A to phase B is 95:5. In some embodiments, the volume ratio of phase A to phase B is 5:95.

[0013] In some embodiments, phase A and phase B are gradient eluted according to the following table:

[0014] Time (min) Phase A (%) Phase B (%) 0 95 5 5 95 5 5.01 5 95 15 5 95

[0015] In some embodiments, the method for separating and detecting 1,3-propane sultone may include or be implemented by the following steps:

[0016] 1) Take an appropriate amount of the sample to be tested containing 1,3-propane sultone, add a solvent to dissolve it, and mix it evenly;

[0017] 2) Set instrument parameters: detector, chromatographic column, ion source, scanning mode, flow rate of mobile phase, column temperature;

[0018] 3) Taking a certain amount of the solution in step 1), injecting it into a high performance liquid chromatography-mass spectrometer to complete the separation and determination of 1,3-propane sultone.

[0019] The solvent described in step 1) is acetonitrile.

[0020] In step 1), each 1 ml of the solvent may contain 5 mg to 40 mg of the sample to be tested. In some embodiments, in step 1), each 1 ml of the solvent may contain 20 mg of the sample to be tested.

[0021] In step 2), the detector is a single quadrupole MS detector.

[0022] In step 2), the ion source may be an ESI ion source.

[0023] In step 2), the scanning mode may be SIM m / z=140(+).

[0024] The flow rate of the mobile phase may be 0.1 ml / min to 0.5 ml / min. In some embodiments, in step 2), the flow rate of the mobile phase is 0.1 ml / min. In some embodiments, in step 2), the flow rate of the mobile phase is 0.3 ml / min. In some embodiments, in step 2), the flow rate of the mobile phase is 0.5 ml / min.

[0025] In step 2), the temperature of the chromatographic column box is 15° C. to 40° C. In some embodiments, the temperature of the chromatographic column box is 20° C.; in some embodiments, the temperature of the chromatographic column box is 35° C.; in some embodiments, the temperature of the chromatographic column box is 40° C.

[0026] In step 3), the sample solution injection volume may be 1 μl to 20 μl. In some embodiments, the sample solution injection volume is 2 μl. In some embodiments, the sample solution injection volume is 3 μl. In some embodiments, the sample solution injection volume is 4 μl.

[0027] In some embodiments, the method for separating and detecting 1,3-propane sultone comprises or can be implemented according to the following steps:

[0028] 1) Take an appropriate amount of the sample containing 1,3-propane sultone, add acetonitrile to dissolve, and mix well;

[0029] 2) Set the flow rate of the mobile phase to 0.1ml / min~0.5ml / min, the detector to a single quadrupole MS detector, the chromatographic column to a Waters Xbridge C18 column, the flow rate of the mobile phase to 0.1ml / min~0.5ml / min, and the column box temperature to 15℃~40℃;

[0030] 3) 1 μl to 20 μl of the sample solution of step 1) is taken and injected into a high performance liquid chromatography-mass spectrometer to complete the separation and determination of 1,3-propane sultone.

[0031] The high performance liquid chromatography-mass spectrometer can be an American Agilent 1260 high performance liquid chromatography-mass spectrometer system and workstation or other suitable and feasible systems.

[0032] In some embodiments, the method for separating and detecting 1,3-propane sultone comprises: using a chromatographic column filled with octadecylsilane bonded phase, the mobile phase is divided into phase A and phase B, phase A is an aqueous solution of alkali, and phase B is acetonitrile.

[0033] In some embodiments, the method for separating and detecting 1,3-propane sultone comprises: using a chromatographic column filled with ethylene bridged hybrid particles (BEH), the mobile phase is divided into phase A and phase B, phase A is 0.1% ammonia solution, and phase B is acetonitrile.

[0034] In some embodiments, the method for separating and detecting 1,3-propane sultone comprises: using a chromatographic column filled with ethylene bridged hybrid particles (BEH), the mobile phase is divided into phase A and phase B, phase A is 0.1% ammonia solution, and phase B is acetonitrile; the phase A and phase B are gradient eluted according to the following table:

[0035] Time (min) Phase A (%) Phase B (%) 0 95 5 5 95 5 5.01 5 95 15 5 95

[0036] By adopting the separation method of the present invention, the time for separating and detecting 1,3-propane sultone can be within 15 minutes.

[0037] In the above or below, regardless of whether the words "about" or "approximately" are used, all the numbers disclosed herein are approximate values. The value of each number may differ by 1%, 2%, 5%, 7%, 8% or 10%.

[0038] The method provided by the present invention can effectively separate the 1,3-propane sultone impurity in the drug, with a separation degree of 1.5 or more, 2.5 or more, or 5 or more, and complete baseline separation, which can solve the problem that the corresponding value of 1,3-propane sultone in the ultraviolet detector is low and the detection result of low-concentration 1,3-propane sultone is inaccurate. The method of the present invention is simple, fast, accurate and highly sensitive. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 Blank solution chromatogram in Example 1;

[0040] Figure 2 Chromatogram of the control solution in Example 1;

[0041] Figure 3 Chromatogram of the test solution in Example 1;

[0042] Figure 4 Chromatogram of the spiked solution of the test sample in Example 1;

[0043] Figure 5 Blank solution chromatogram in Example 2;

[0044] Figure 6 Chromatogram of the control solution in Example 2;

[0045] Figure 7 Chromatogram of the test solution in Example 2;

[0046] Figure 8 Chromatogram of the spiked solution of the test sample in Example 2;

[0047] Fig. 9 Blank solution chromatogram in Example 3;

[0048] Fig.10 Chromatogram of detection limit solution 1 in Example 3;

[0049] Fig.11 Chromatogram of detection limit solution 2 in Example 3;

[0050] Fig.12 Chromatogram of detection limit solution 3 in Example 3;

[0051] Fig.13 Chromatogram of the control solution in Example 3;

[0052] Fig.14 Chromatogram of the test solution in Example 3;

[0053] Fig.15 Chromatogram of test sample spiked solution 1 in Example 3;

[0054] Fig.16 Chromatogram of test sample spiked solution 2 in Example 3;

[0055] Fig.17 Chromatogram of test sample spiked solution 3 in Example 3. DETAILED DESCRIPTION

[0056] The embodiment of the present invention discloses a method for separating and detecting 1,3-propane sultone. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It is particularly important to point out that all similar substitutions and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The method of the present invention has been described through a preferred embodiment, and relevant personnel can obviously modify or appropriately change and combine the methods described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0057] In the embodiment of the present invention,

[0058] The content of residual 1,3-propane sultone in the test sample was calculated according to the following formula:

[0059]

[0060] Where: A T is the peak area of ​​the impurity to be detected in the test solution;

[0061] A S The average value of the peak area of ​​the impurity to be tested by three consecutive injections of the reference solution;

[0062] W T is the sample weight of the test solution, mg;

[0063] W S The mass of the impurity to be tested weighed to prepare the reference solution, mg;

[0064] D T is the dilution factor of the test solution;

[0065] D S is the dilution multiple of the reference solution;

[0066] P is the content of the reference substance, %.

[0067] The recovery rate is calculated as follows:

[0068]

[0069] Determination of added amount (μg) = W T+S ×(C T+S -C T )×1000,

[0070] Theoretical addition amount (μg) = C S ×V,

[0071]

[0072] Where: W T+S is the sample amount of the spiked solution of the test sample, mg;

[0073] C T+S is the impurity residue measured in the spiked solution of the test sample, ppm;

[0074] C T is the residual amount of impurities measured in the test solution, ppm;

[0075] C S is the concentration of impurities in the reference stock solution added to the test sample spike solution, μg / ml;

[0076] V is the volume of the reference stock solution added to the test sample spike solution, ml;

[0077] W S is the weight of the corresponding impurity in the reference stock solution, mg;

[0078] D S is the dilution multiple of the reference solution;

[0079] P is the content of the reference substance, %.

[0080] In the present invention, N / A means none.

[0081] In order to further understand the present invention, the present invention is described in detail below in conjunction with embodiments.

[0082] Instrument: American Agilent 1260 high performance liquid chromatography-mass spectrometry system and workstation; automatic injection.

[0083] Example 1

[0084] Instruments and conditions

[0085] Chromatographic conditions:

[0086] Chromatographic column: Waters Xbridge C18 column, 2.1*150mm, 3.5μm;

[0087] Detector: Single quadrupole MS;

[0088] Ion source: ESI;

[0089] Scan mode: SIM m / z = 140 (+);

[0090] Flow rate: 0.3ml / min;

[0091] Column temperature: 35°C;

[0092] Injection volume: 2 μl;

[0093] Mobile phase A: 0.1% NH3.H2O

[0094] Mobile phase B: acetonitrile;

[0095] Elution gradient:

[0096] Time (min) Phase A Phase B 0 90 5 5 90 5 5.01 10 90 15 10 90

[0097] Running time: 15min;

[0098] Post-running time: 5min;

[0099] Experimental procedures

[0100] Blank solution: acetonitrile.

[0101] 1,3-Propane sultone reference substance stock solution 1: Weigh 62.70 mg of 1,3-propane sultone into a 25 ml volumetric flask and dissolve it to volume with acetonitrile.

[0102] 1,3-Propane sultone reference stock solution 2: Pipette 1.0 ml of reference stock solution 1 into a 50 ml volumetric flask and dilute to volume with acetonitrile.

[0103] 1,3-Propane sultone reference solution: Pipette 1.0 ml of reference stock solution 2 into a 50 ml volumetric flask and dilute to volume with acetonitrile.

[0104] Test solution: weigh about 100 mg of the test sample volapivir into a 5 ml volumetric flask and dissolve it to volume with acetonitrile.

[0105] Test sample spiked solution: weigh about 100 mg of the test sample volapivir into a 5 ml volumetric flask and dissolve it to volume with 1,3-propane sultone reference solution.

[0106] Take blank solution, reference solution, test solution, and test solution spiked with the same conditions and analyze them. Record the chromatogram. The results are shown in Table 1 below:

[0107] Table 1:

[0108]

[0109] Example 2

[0110] Instruments and conditions

[0111] Chromatographic conditions:

[0112] Chromatographic column: Waters Xbridge C18 column, 2.1*150mm, 3.5μm;

[0113] Detector: Single quadrupole MS;

[0114] Ion source: ESI;

[0115] Scan mode: SIM m / z = 140 (+);

[0116] Flow rate: 0.3ml / min;

[0117] Column temperature: 35°C;

[0118] Injection volume: 4 μl;

[0119] Mobile phase A: 0.1% NH3.H2O;

[0120] Mobile phase B: acetonitrile;

[0121] Elution gradient:

[0122]

[0123]

[0124] Running time: 15min;

[0125] Post-run: 5min;

[0126] Experimental procedures

[0127] Blank solution: acetonitrile;

[0128] 1,3-Propane sultone reference stock solution 1: Weigh 62.70 mg of 1,3-propane sultone into a 25 ml volumetric flask and dissolve it in acetonitrile to make up to volume;

[0129] 1,3-Propane sultone reference stock solution 2: Pipette 1.0 ml of reference stock solution 1 into a 50 ml volumetric flask and dilute to volume with acetonitrile;

[0130] 1,3-Propane sultone reference solution: Pipette 1.0 ml of reference stock solution 2 into a 50 ml volumetric flask and dilute to volume with acetonitrile;

[0131] Test solution: weigh about 100 mg of the test substance volapivir into a 5 ml volumetric flask and dissolve it to volume with acetonitrile;

[0132] Test sample spike solution: weigh about 100 mg of the test sample volapivir into a 5 ml volumetric flask and dissolve it to volume with 1,3-propane sultone reference solution;

[0133] Take blank solution, reference solution, test solution, and test solution spiked with the same solution and perform LC-MS analysis according to the above conditions. Record the chromatogram. The results are shown in Table 2 below:

[0134] Table 2:

[0135]

[0136] Example 3

[0137] Instruments and conditions

[0138] Chromatographic conditions:

[0139] Chromatographic column: Waters Xbridge C18 column, 2.1*150mm, 3.5μm;

[0140] Detector: Single quadrupole MS;

[0141] Ion source: ESI;

[0142] Scan mode: SIM m / z = 140 (+);

[0143] Flow rate: 0.3ml / min;

[0144] Column temperature: 35°C;

[0145] Injection volume: 4 μl;

[0146] Mobile phase A: 0.1% NH3.H2O;

[0147] Mobile phase B: acetonitrile;

[0148] Elution gradient:

[0149] Time (min) Phase A Phase B 0 95 5 5 95 5 5.01 5 95 15 5 95

[0150] Running time: 15min;

[0151] Post-running: 5min;

[0152] Experimental procedures

[0153] Blank solution: acetonitrile.

[0154] 1,3-Propane sultone reference substance stock solution 1: Weigh 62.25 mg of 1,3-propane sultone into a 25 ml volumetric flask and dissolve it to volume with acetonitrile.

[0155] 1,3-Propane sultone reference stock solution 2: Pipette 1.0 ml of reference stock solution 1 into a 50 ml volumetric flask and dilute to volume with acetonitrile.

[0156] 1,3-Propane sultone reference solution: Pipette 1.0 ml of reference stock solution 2 into a 50 ml volumetric flask and dilute to volume with acetonitrile.

[0157] Test solution: weigh about 100 mg of the test sample volapivir into a 5 ml volumetric flask and dissolve it to volume with acetonitrile.

[0158] Test sample spiked solution: weigh about 100 mg of the test sample volapivir into a 5 ml volumetric flask and dissolve it to volume with 1,3-propane sultone reference solution. Prepare three replicates in parallel.

[0159] Detection limit solution: Pipette 3.0 ml of 1,3-propane sultone reference solution into a 10 ml volumetric flask and dilute to volume with acetonitrile.

[0160] Take blank solution, detection limit solution, reference solution, test solution, and test spiked solution and analyze them according to the following sequence table (Table A) and the above conditions, record the chromatograms, and the results are shown in Tables 3 to 7.

[0161] Table A:

[0162]

[0163]

[0164] Result analysis:

[0165] Table 3: System suitability results

[0166]

[0167] Table 4: Specificity results

[0168]

[0169]

[0170] Table 5: Detection limit results

[0171]

[0172] Table 6: Accuracy results

[0173]

[0174] Table 7: Solution stability results

[0175]

[0176]

[0177] It can be seen from Example 3 that the method of the present invention has the advantages of being simple, rapid, highly sensitive and highly accurate in detecting the impurity 1,3-propane sultone in drugs.

Claims

1. A method for separating and detecting 1,3-propane sultone, characterized in that: include: A chromatographic column using an octadecylsilane bonded phase as filler, wherein the mobile phase is divided into a phase A and a phase B, wherein the phase A is an aqueous solution of ammonia or an ammonium salt, and the phase B is acetonitrile; the chromatographic column filler is ethylene bridge hybrid particles; and the method comprises the following steps: 1) Take an appropriate amount of the sample to be tested containing 1,3-propane sultone, add a solvent to dissolve it, and mix it evenly; 2) Set instrument parameters: detector, chromatographic column, ion source, scanning mode, flow rate of mobile phase, column temperature; 3) taking a certain amount of the solution of step 1) and injecting it into a high performance liquid chromatography-mass spectrometer to complete the separation and determination of 1,3-propane sultone; wherein the flow rate of the mobile phase is 0.1 ml / min to 0.5 ml / min; the detector is a single quadrupole MS detector; the scanning mode is SIM m / z=140(+); the ion source is an ESI ion source; and the column box temperature of the chromatographic column is 15°C to 40°C.

2. The method according to claim 1, wherein the chromatographic column is selected from the group consisting of Waters Xbridge C18 column and Waters XBridge TM C 18 A type of column.

3. according to the arbitrary described method of claim 1-2, described ammonium salt is selected from one among ammonium formate and ammonium acetate.

4. The method according to claim 1, wherein the volume ratio of ammonia water or ammonium salt to water in phase A is 5:10000 to 2:1000.

5. The method according to claim 1, wherein the volume ratio of phase A to phase B is 95:5 to 5:

95.

6. The method according to claim 1, wherein the solvent is acetonitrile; and / or each 1 ml of the solvent contains 5 mg to 40 mg of the sample to be tested.

7. The method according to claim 1, wherein: Step 3) The sample solution injected is 1 μl to 20 μl.

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