A method for measuring the chromatographic purity of 2,4,6-trimethylbenzenesulfonyl chloride using HPLC

Abstract

The present invention relates to the technical field of detection and analysis, and in particular to a method for determining the chromatographic purity of 2,4,6-trimethylbenzenesulfonyl chloride by utilizing HPLC. The method comprises the following conditions: a stationary phase is an octadecylsilane bonded phase, a mobile phase comprises a mobile phase A and a mobile phase B, the mobile phase A is a potassium dihydrogen phosphate buffered saline solution, and the mobile phase B is acetonitrile; the method can effectively separate 2,4,6-trimethylbenzenesulfonyl chloride, mesitylene, and a hydrolysis impurity 2,4,6-trimethylbenzenesulfonic acid in a 2,4,6-trimethylbenzenesulfonyl chloride product, has high separation degree and high sensitivity, does not generate interfering impurities, and can accurately determine the purity of the 2,4,6-trimethylbenzenesulfonyl chloride in the product.

Description

Technical Field

[0001] The present invention relates to the technical field of detection and analysis, in particular to a method for measuring the chromatographic purity of 2,4,6-trimethylbenzenesulfonyl chloride by utilizing HPLC. Background Art

[0002] 2,4,6-Trimethylbenzenesulfonyl chloride is an organic compound synthesized by the reaction of trimethylbenzene with chlorosulfonic acid. Its molecular formula is C9H11ClO2S, and its molecular structure contains a sulfonyl group and three methyl groups. 2,4,6-Trimethylbenzenesulfonyl chloride is often used as a catalyst, catalyzing C-H bond activation, Heck reactions, and Suzuki coupling reactions. Furthermore, 2,4,6-Trimethylbenzenesulfonyl chloride is an important biochemical and pharmaceutical reagent, serving as a condensation reagent in nucleoside synthesis. It can also be used in intramolecular esterification, amino acid cyclization, guanidine protecting groups, and the preparation of biologically active compounds such as sulfonamide compounds.

[0003] In the prior art, 2,4,6-trimethylbenzenesulfonyl chloride is mainly produced by the reaction of trimethylbenzene and chlorosulfonic acid. In order to control the quality of the obtained 2,4,6-trimethylbenzenesulfonyl chloride product, its purity needs to be tested. The commonly used method is high-performance liquid chromatography (HPLC). The HPLC method in the prior art usually contains reagents such as water and methanol. 2,4,6-trimethylbenzenesulfonyl chloride is sensitive to water and will hydrolyze to form the hydrolysis impurity 2,4,6-trimethylbenzenesulfonic acid. At the same time, methanol, as a derivatization reagent, will also produce hydrolysis impurities, resulting in the inability to accurately detect the quality of 2,4,6-trimethylbenzenesulfonyl chloride. Summary of the Invention

[0004] In order to overcome the problem in the prior art of using HPLC to test 2,4,6-trimethylbenzenesulfonyl chloride that 2,4,6-trimethylbenzenesulfonyl chloride produces hydrolysis impurities and methanol hydrolysis impurities, which makes it impossible to accurately detect the quality of 2,4,6-trimethylbenzenesulfonyl chloride, the present application provides a method for determining the chromatographic purity of 2,4,6-trimethylbenzenesulfonyl chloride using HPLC. The method has the technical effects of accurately determining the chromatographic purity of 2,4,6-trimethylbenzenesulfonyl chloride, and simultaneously detecting the raw material mesitylene and the hydrolysis impurity 2,4,6-trimethylbenzenesulfonic acid, with high separation and high sensitivity.

[0005] The specific technical solutions of the present invention are:

[0006] A method for determining the chromatographic purity of 2,4,6-trimethylbenzenesulfonyl chloride by using HPLC, comprising the following conditions: the stationary phase is an octadecylsilane bonded phase, the mobile phase comprises a mobile phase A and a mobile phase B, the mobile phase A is a potassium dihydrogen phosphate buffered saline solution, and the mobile phase B is acetonitrile.

[0007] Preferably, the method comprises the following steps:

[0008] Step 1: Prepare the sample to be tested;

[0009] Step 2: injecting the sample to be tested in step 1 into a liquid chromatography device for gradient elution and determining the liquid chromatography purity of the sample to be tested;

[0010] The gradient elution includes: a first gradient, a second gradient, a third gradient, a fourth gradient and a fifth gradient;

[0011] The gradient elution conditions include: a flow rate of 1.0 ml / min, a chromatographic column temperature of 30° C., a mobile phase comprising: a mobile phase A with a volume fraction of 25-90% and a mobile phase B with a volume fraction of 10-75%, and an octadecylsilane bonded phase as the stationary phase.

[0012] Preferably, the conditions of the first gradient include: switching the first gradient mobile phase at the 0th minute, the first gradient mobile phase including mobile phase A with a volume fraction of 90% and mobile phase B with a volume fraction of 10%.

[0013] Preferably, the conditions of the second gradient include: switching the second gradient mobile phase at 8 minutes, the second gradient mobile phase including mobile phase A with a volume fraction of 40% and mobile phase B with a volume fraction of 60%.

[0014] Preferably, the conditions of the third gradient include: switching the third gradient mobile phase at the 25th minute, the third mobile phase including mobile phase A with a volume fraction of 25% and mobile phase B with a volume fraction of 75%.

[0015] Preferably, the fourth gradient condition includes: switching the fourth gradient mobile phase at 25.1 min, the fourth gradient mobile phase including mobile phase A with a volume fraction of 90% and mobile phase B with a volume fraction of 10%.

[0016] Preferably, the conditions of the fifth gradient include: switching the fifth gradient mobile phase at 30 minutes, the fifth gradient mobile phase including mobile phase A with a volume fraction of 90% and mobile phase B with a volume fraction of 10%.

[0017] The present application provides a method for determining the chromatographic purity of 2,4,6-trimethylbenzenesulfonyl chloride using HPLC. The method has the technical effects of accurately determining the chromatographic purity of 2,4,6-trimethylbenzenesulfonyl chloride and simultaneously detecting the raw material mesitylene and the hydrolyzed impurity 2,4,6-trimethylbenzenesulfonic acid with high separation and high sensitivity. The method uses HPLC to determine the trimethylbenzenesulfonyl chloride product, and can accurately determine the chromatographic purity of 2,4,6-trimethylbenzenesulfonyl chloride, mesitylene, and hydrolyzed impurity 2,4,6-trimethylbenzenesulfonic acid. The stationary phase in the HPLC method uses an octadecylsilane bonded phase, and the mobile phases use mobile phase A and mobile phase B. Mobile phase A is a potassium dihydrogen phosphate buffer solution, and mobile phase B is acetonitrile. The method uses a gradient elution method, and the ratio of mobile phase A to mobile phase B is adjusted under different gradients. The method can effectively separate 2,4,6-trimethylbenzenesulfonyl chloride, mesitylene, and hydrolyzed impurity 2,4,6-trimethylbenzenesulfonic acid in the 2,4,6-trimethylbenzenesulfonyl chloride product, has high separation and sensitivity, does not produce interfering impurities, and can accurately determine the purity of 2,4,6-trimethylbenzenesulfonyl chloride in the product.

[0018] Preferably, the step of preparing the sample to be tested in step 1 includes: weighing 2,4,6-trimethylbenzenesulfonyl chloride to be tested and placing it in a volumetric flask, adding acetonitrile and aniline, and then using acetonitrile to adjust the volume to obtain the sample to be tested.

[0019] Preferably, the liquid chromatography purity of the sample to be tested includes one or more of 2,4,6-trimethylbenzenesulfonic acid, 2,4,6-trimethylbenzenesulfonyl chloride and trimethylbenzene.

[0020] Preferably, the detection wavelength of the liquid chromatography device is 210 nm.

[0021] Compared with the existing technology, this application has the following technical effects:

[0022] It can effectively separate 2,4,6-trimethylbenzenesulfonyl chloride, mesitylene, and hydrolysis impurity 2,4,6-trimethylbenzenesulfonic acid in the 2,4,6-trimethylbenzenesulfonyl chloride product with high separation and sensitivity, without producing interfering impurities, and can accurately determine the purity of 2,4,6-trimethylbenzenesulfonyl chloride in the product. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is the HPLC chromatogram of Example 1 of the present application, where b is the HPLC chromatogram of the sample to be tested, and a is the HPLC chromatogram of the aniline-derived blank.

[0024] Figure 2This is the HPLC chromatogram of Comparative Example 1 of the present application, where a is the HPLC chromatogram of the sample to be tested, and b is the control HPLC chromatogram of a mixed solution of mesitylene, 2,4,6-trimethylbenzenesulfonic acid and 2,4,6-trimethylbenzenesulfonyl chloride derivatives. DETAILED DESCRIPTION

[0025] The present invention will be further described below with reference to the embodiments.

[0026] Example 1:

[0027] A method for determining the chromatographic purity of 2,4,6-trimethylbenzenesulfonyl chloride using HPLC comprises the following steps:

[0028] Step 1: Prepare the sample to be tested;

[0029] Step 2: injecting the sample to be tested in step 1 into a liquid chromatography device for gradient elution and determining the liquid chromatography purity of the sample to be tested, the gradient elution including: a first gradient, a second gradient, a third gradient, a fourth gradient and a fifth gradient; switching the first gradient mobile phase at 0 min, the first gradient mobile phase including a mobile phase A with a volume fraction of 90% and a mobile phase B with a volume fraction of 10%, switching the second gradient mobile phase at 8 min, the second gradient mobile phase including a mobile phase A with a volume fraction of 40% and a mobile phase B with a volume fraction of 60%, switching the third gradient mobile phase at 25 min, the third mobile phase including a mobile phase A with a volume fraction of 25% and a mobile phase B with a volume fraction of 75%, switching the fourth gradient mobile phase at 25.1 min, the fourth gradient mobile phase including a mobile phase A with a volume fraction of 90% and a mobile phase B with a volume fraction of 10%, switching the fifth gradient mobile phase at 30 min, the fifth gradient mobile phase including a mobile phase A with a volume fraction of 90% and a mobile phase B with a volume fraction of 10%.

[0030] The preparation of the sample to be tested includes the following steps: taking about 20 mg of the 2,4,6-trimethylbenzenesulfonyl chloride sample to be tested, accurately weighing it, placing it in a 100 mL volumetric flask, adding 10.0 mL of acetonitrile, then adding 1.0 mL of aniline, ultrasonicating for 3 minutes, standing for 2 hours, diluting it to the scale with acetonitrile, and shaking it well. The injection volume of the test solution is about 5 μL;

[0031] The liquid chromatography apparatus was an Agilent 1100 high performance liquid chromatography system and workstation from the United States, and the detector was a DAD detector (detection wavelength 210 nm).

[0032] The stationary phase was an Agilent ZORBAX Eclipse XDB-C18 column (4.6×150 mm, 3.5 μm), the mobile phase A was potassium dihydrogen phosphate buffer solution (0.01 mol / L), and the mobile phase B was acetonitrile. The flow rate was 1.0 ml / min, and the column temperature was 30°C.

[0033] Elution conditions are shown in Table 1.

[0034] Table 1. Elution gradient conditions of Example 1

[0035]

[0036] Comparative Example 1:

[0037] A method for determining the chromatographic purity of 2,4,6-trimethylbenzenesulfonyl chloride using HPLC comprises the following steps:

[0038] Step 1: Prepare the sample to be tested;

[0039] Step 2: injecting the sample to be tested in step 1 into a liquid chromatography device for gradient elution and determining the liquid chromatography purity of the sample to be tested, wherein the gradient elution includes: a first gradient, a second gradient, a third gradient, a fourth gradient and a fifth gradient;

[0040] The preparation of the sample to be tested includes the following steps: taking about 20 mg of the 2,4,6-trimethylbenzenesulfonyl chloride sample to be tested, accurately weighing it, placing it in a 100 mL volumetric flask, diluting it to the scale with acetonitrile, and shaking it well. The injection volume of the test solution is about 5 μL;

[0041] The liquid chromatography apparatus was an Agilent 1100 high performance liquid chromatography system and workstation from the United States, and the detector was a DAD detector (detection wavelength 210 nm).

[0042] The stationary phase was an Agilent ZORBAX Eclipse XDB-C18 column (4.6×150 mm, 3.5 μm), the mobile phase A was potassium dihydrogen phosphate buffer solution (0.01 mol / L), and the mobile phase B was acetonitrile. The flow rate was 1.0 ml / min, and the column temperature was 30°C.

[0043] Elution conditions are shown in Table 2.

[0044] Table 2, elution gradient conditions of Comparative Example 1

[0045]

[0046] like Figure 1As shown, aniline was added to the test sample of Example 1. The results showed that the retention time of the derivative product of 2,4,6-trimethylbenzenesulfonyl chloride was 12.685 min, no hydrolysis impurities were generated at 5.035 min, and it was well separated from the raw material mesitylene and the hydrolysis impurity 2,4,6-trimethylbenzenesulfonic acid. The signal-to-noise ratio at the LOQ level (concentration: 0.00005 mg / mL) was 11.9, with high sensitivity. The test solution after derivatization was stable at room temperature within 24 h. This method is stable and reliable. Figure 2 As shown. In Comparative Example 1, aniline was not added to the test sample. The results showed that the retention time of 2,4,6-trimethylbenzenesulfonyl chloride was 17.155 min, and 2.6% of hydrolysis impurities were generated at 5.035 min. The chromatographic purity of 2,4,6-trimethylbenzenesulfonyl chloride could not be accurately detected, and the method was not applicable. The above results show that the method provided by the present application can accurately determine the purity of 2,4,6-trimethylbenzenesulfonyl chloride, and can also detect mesitylene and hydrolysis impurity 2,4,6-trimethylbenzenesulfonic acid, and can efficiently separate 2,4,6-trimethylbenzenesulfonyl chloride, mesitylene and hydrolysis impurity 2,4,6-trimethylbenzenesulfonic acid with high separation degree and high accuracy.

[0047] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any way. Any simple modification, change and equivalent transformation made to the above embodiment based on the technical essence of the present invention still fall within the scope of protection of the technical solution of the present invention.

Claims

1. A method for determining the chromatographic purity of 2,4,6-trimethylbenzenesulfonyl chloride using HPLC, characterized in that: The following steps are involved: Step 1: Weigh 2,4,6-trimethylbenzenesulfonyl chloride into a volumetric flask, add acetonitrile and aniline, and then dilute to volume with acetonitrile to obtain the sample to be tested; Step 2: injecting the sample to be tested in step 1 into a liquid chromatography device for gradient elution and determining the liquid chromatography purity of the sample to be tested, the detection wavelength of the liquid chromatography device is 210 nm; The gradient elution includes: a first gradient, a second gradient, a third gradient, a fourth gradient and a fifth gradient; Mobile phase A is potassium dihydrogen phosphate buffered saline solution, mobile phase B is acetonitrile, and the stationary phase is octadecylsilane bonded phase; the gradient elution conditions are: 。 2. The method according to claim 1, wherein: The volume of acetonitrile in step 1 was 10.0 ml.

3. The method according to claim 1, wherein: The volume of aniline in step 1 was 1.0 mL.

4. The method according to claim 1, wherein: The volume of the volumetric flask in step 1 is 100 mL.

5. The method according to claim 1, wherein: The injection volume of the sample to be tested in step 2 is 5 μL.

6. The method according to claim 1, wherein: The column temperature of the liquid chromatography device in step 2 is 30°C.

7. The method according to claim 1, wherein: The flow rate of mobile phase B in step 2 was 1.0 ml / min.

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