Method for detecting impurity content in etravirine intermediate 1
阅读说明：本技术 一种依曲韦林中间体1中杂质含量检测方法 (Method for detecting impurity content in etravirine intermediate 1 ) 是由 汪士金 范康梅 孙少发 王国华 谭梓骏 于 2021-03-03 设计创作，主要内容包括：本发明涉及药物分析技术领域,且公开了一种依曲韦林中间体1中杂质含量检测方法,包括以下步骤,步骤一：将依曲韦林中间体1以十八烷基硅烷键合硅烷为填充剂的色谱柱；步骤二：以有机相与水相作为流动相梯度洗脱；步骤三：进行高效液相色谱法分析测定,测定依曲韦林中间体1及其六个杂质的含量。该依曲韦林中间体1中杂质含量检测方法,该方法工艺简单、成本低廉,采用梯度洗脱的方法,能够测定依曲韦林中间体1杂质的含量,检测方法科学、合理、客观,从而能够更好地控制依曲韦林的质量。(The invention relates to the technical field of pharmaceutical analysis and discloses a method for detecting the content of impurities in an etravirine intermediate 1, which comprises the following steps: a chromatographic column using the etravirine intermediate 1 and octadecylsilane chemically bonded silane as a filler; step two: taking an organic phase and a water phase as a mobile phase for gradient elution; step three: and (4) carrying out high performance liquid chromatography analysis and determination to determine the contents of the etravirine intermediate 1 and six impurities thereof. The method for detecting the impurity content in the etravirine intermediate 1 has the advantages of simple process and low cost, can determine the impurity content of the etravirine intermediate 1 by adopting a gradient elution method, and is scientific, reasonable and objective, so that the quality of etravirine can be better controlled.)
1. A method for detecting the content of impurities in an etravirine intermediate 1 is characterized by comprising the following steps,
the method comprises the following steps: a chromatographic column using the etravirine intermediate 1 and octadecylsilane chemically bonded silane as a filler;
step two: taking an organic phase and a water phase as a mobile phase for gradient elution;
step three: and (4) carrying out high performance liquid chromatography analysis and determination to determine the contents of the etravirine intermediate 1 and six impurities thereof.
2. The method for detecting the content of impurities in the etravirine intermediate 1 according to the step two of claim 1, which is characterized in that: the organic phase is acetonitrile, and the aqueous phase is formate.
3. The method for detecting the content of impurities in the etravirine intermediate 1 according to the step two of claim 1, which is characterized in that: the volume ratio of the buffer salt solution to the acetonitrile in the mobile phase is 70-10: 30-90, and preferably 70: 30.
4. The method for detecting the content of impurities in the etravirine intermediate 1 according to the step two of claim 1, which is characterized in that: the concentration of the aqueous formate solution is 0.01mol/L ammonium formate and 0.2% formic acid.
5. The method for detecting the content of impurities in the etravirine intermediate 1 according to the step two of claim 1, which is characterized in that: the detection wavelength is 254nm, and the column temperature is preferably 35 ℃.
6. The method for detecting the content of impurities in the etravirine intermediate 1 according to the step two of claim 1, which is characterized in that:
the gradient wash conditions were:
。
7. The method for detecting the content of impurities in the etravirine intermediate 1 according to the third step of claim 1, wherein the method comprises the following steps:
the method for measuring the content of the intermediate 1 of the trinetrvirin and six impurities of the intermediate comprises the following steps,
a. taking a proper amount of an etravirine intermediate 1 sample, dissolving the sample with acetonitrile or formate, and preparing a sample solution containing 10.1-0.6 mg of etravirine intermediate per 1 mL;
b. setting the flow rate of a mobile phase at 101mL/min, the detection wavelength at 254nm and the column temperature at 35 ℃;
c. and (c) injecting 10 mu L of the sample solution of the step (a) into a high performance liquid chromatograph to complete content determination.
8. The method for detecting the content of impurities in the etravirine intermediate 1 according to the third step of claim 1, wherein the method comprises the following steps: the six impurities are respectively: impurity 1: 2,4, 6-trichloropyrimidine; impurity 2: 3, 5-dimethyl-4-hydroxybenzonitrile; impurity 3: 4- ((4, 6-dichloropyrimidine-2-benzyl) oxy) -3, 5-dimethylbenzonitrile; impurity 4: 4,4' - ((6-chloropyrimidine-2, 4) -benzyloxy) - (3, 5-benzonitrile); impurity 5: 4-hydroxy-3, 5-dimethylbenzoic acid; impurity 6: 4- ((2, 6-dichloropyrimidine-4-benzyl) oxy) -3, 5-dimethylbenzoic acid.
Technical Field
The invention relates to the technical field of pharmaceutical analysis, in particular to a method for detecting the content of impurities in an etravirine intermediate 1.
Background
1, 4- ((2, 6-dichloropyrimidine-4-benzyl) oxy) -3, 5-dimethyl benzonitrile serving as an etravirine intermediate has a molecular formula of C13H9N3Cl2O and a molecular weight of 294,
the structural formula is as follows:,
etravirine can be used in combination with other antiretroviral drugs in adult HIV-1 infected patients who develop resistance after initial treatment with antiretroviral drugs.
Etravirine belongs to the non-nucleoside reverse transcriptase inhibitor (NNRTI) of human immunodeficiency virus type 1 (HIV-1), can be directly combined with HIV-1 reverse transcriptase, and can block the activities of RNA-dependent and DNA-dependent DNA polymerase by destroying the catalytic site of enzyme, and the etravirine can not inhibit human alpha, beta and gamma type DNA polymerase.
In the prior art, the method for detecting the impurity content in the etravirine intermediate 1 is difficult to determine the impurity content of the etravirine intermediate 1, cannot carry out scientific, reasonable and objective detection, and is difficult to better control the quality of etravirine, so that the method for detecting the impurity content in the etravirine intermediate 1 is provided.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides the method for detecting the impurity content in the etravirine intermediate 1, which can be used for detecting the impurity content of the etravirine intermediate 1, is scientific, reasonable and objective, can better control the quality of etravirine and the like, and solves the problems that the impurity content in the etravirine intermediate 1 is difficult to detect, the impurity content of the etravirine intermediate 1 cannot be scientifically, reasonably and objectively detected, and the quality of the etravirine 1 cannot be better controlled in the existing method for detecting the impurity content in the etravirine intermediate 1 in the prior art.
In order to realize the purpose that the content of the impurities of the etravirine intermediate 1 can be measured, and the detection method is scientific, reasonable and objective, so that the quality of etravirine can be better controlled, the invention provides the following technical scheme: a method for detecting the content of impurities in an etravirine intermediate 1 comprises the following steps,
the method comprises the following steps: a chromatographic column using the etravirine intermediate 1 and octadecylsilane chemically bonded silane as a filler;
step two: taking an organic phase and a water phase as a mobile phase for gradient elution;
step three: and (4) carrying out high performance liquid chromatography analysis and determination to determine the contents of the etravirine intermediate 1 and six impurities thereof.
Preferably, the organic phase is acetonitrile and the aqueous phase is formate.
Preferably, the volume ratio of the buffer salt solution to the acetonitrile in the mobile phase is 70-10: 30-90, and preferably 70: 30.
Preferably, the concentration of the aqueous formate solution is 0.01mol/L ammonium formate and 0.2% formic acid.
Preferably, the detection wavelength is 254nm, and the column temperature is preferably 35 ℃.
Preferably, the gradient wash conditions are:
time (minutes)
Water phase: 0.01mol/L ammonium formate and 0.2% formic acid
Organic phase: acetonitrile
0
70
30
3
70
30
40
10
90
45
10
90
45.1
70
30
55
70
30
Wherein, the method for measuring the content of the intermediate 1 of the trinetrvirin and six impurities of the intermediate comprises the following steps,
a. taking a proper amount of an etravirine intermediate 1 sample, dissolving the sample with acetonitrile or formate, and preparing a sample solution containing 10.1-0.6 mg of etravirine intermediate per 1 mL;
b. setting the flow rate of a mobile phase at 101mL/min, the detection wavelength at 254nm and the column temperature at 35 ℃;
c. and (c) injecting 10 mu L of the sample solution of the step (a) into a high performance liquid chromatograph to complete content determination.
Wherein, the six impurities are respectively: impurity 1: 2,4, 6-trichloropyrimidine; impurity 2: 3, 5-dimethyl-4-hydroxybenzonitrile; impurity 3: 4- ((4, 6-dichloropyrimidine-2-benzyl) oxy) -3, 5-dimethylbenzonitrile; impurity 4: 4,4' - ((6-chloropyrimidine-2, 4) -benzyloxy) - (3, 5-benzonitrile); impurity 5: 4-hydroxy-3, 5-dimethylbenzoic acid; impurity 6: 4- ((2, 6-dichloropyrimidine-4-benzyl) oxy) -3, 5-dimethylbenzoic acid.
Compared with the prior art, the invention provides a method for detecting the content of impurities in an etravirine intermediate 1, which has the following beneficial effects:
the method for detecting the impurity content in the etravirine intermediate 1 has the advantages that an octadecyl silane bonded silane is used as a chromatographic column of a filling agent, an organic phase and a water phase are used as mobile phases for gradient elution, and the impurity content of the etravirine intermediate 1 is measured.
Drawings
FIG. 1 is a schematic representation of a typical map of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Referring to fig. 1, a method for detecting the content of impurities in an etravirine intermediate 1 comprises the following steps,
the method comprises the following steps: a chromatographic column using the etravirine intermediate 1 and octadecylsilane chemically bonded silane as a filler;
step two: taking an organic phase and a water phase as a mobile phase for gradient elution;
step three: performing high performance liquid chromatography analysis and determination to determine the contents of the etravirine intermediate 1 and six impurities thereof;
wherein, the steps for measuring the contents of the etravirine intermediate 1 and six impurities thereof are as follows,
taking a proper amount of an etravirine intermediate 1 sample, dissolving the sample with acetonitrile or formate, and preparing a sample solution containing 10.1-0.6 mg of etravirine intermediate per 1 mL; setting the flow rate of a mobile phase at 101mL/min, the detection wavelength at 254nm and the column temperature at 35 ℃; and (c) injecting 10 mu L of the sample solution of the step (a) into a high performance liquid chromatograph to complete content determination.
Wherein, the gradient washing conditions are as follows:
time (minutes)
Water phase: 0.01mol/L ammonium formate and 0.2% formic acid
Organic phase: acetonitrile
0
70
30
3
70
30
40
10
90
45
10
90
45.1
70
30
55
70
30
The method for detecting the impurity content in the etravirine intermediate 1 has the advantages that an octadecyl silane bonded silane is used as a chromatographic column of a filling agent, an organic phase and a water phase are used as mobile phases for gradient elution, and the impurity content of the etravirine intermediate 1 is measured.
It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.