阅读说明：本技术 一种沙坦类药物合成中杂质的检测方法 (Method for detecting impurities in sartan drug synthesis ) 是由 汤伟彬 蔡强 刘杰 陆国琦 李达胜 兰柳琴 于 2021-01-21 设计创作，主要内容包括：本发明属于药物分析技术领域,具体涉及一种沙坦类药物合成中杂质的检测方法,该方法使用方便快捷的LCMS/MS,对叠氮沙坦联苯或叠氮代沙坦联苯四氮唑进行检测,可用于监测沙坦类药物的质量。本发明首次提出了叠氮沙坦联苯或叠氮代沙坦联苯四氮唑的检测方法,具有准确度高、精密度高、重现性好、稳定性好、专属性强等特点,同时本发明的测定方法操作简单,所用时间比较短,成本低,耗时短。(The invention belongs to the technical field of drug analysis, and particularly relates to a method for detecting impurities in sartan drug synthesis. The invention provides a method for detecting azides sartanbiphenyl or azides sartanbiphenyl tetrazole for the first time, and the method has the characteristics of high accuracy, high precision, good reproducibility, good stability, strong specificity and the like.)

1. A method for detecting impurities in the synthesis of sartan drugs, wherein the structural formula of the impurities is as follows:wherein R is CN orAny one of the above, the detection method comprising the steps of:

(1) preparing solutions, namely respectively preparing a diluent, a blank solution, a reference substance solution and a test solution, wherein the reference substance solution comprises an impurity stock solution and a reference substance stock solution;

(2) the determination method comprises the following steps: measuring the content of impurities by adopting LC-MS/MS, respectively adding a blank solution, a reference solution and a test solution after the system is stable, and recording spectrograms;

i: the chromatographic conditions are as follows: octadecylsilane chemically bonded silica is used as a filling agent, the flow rate is 0.5mL/min +/-0.1 mL/min, the column temperature is as follows: 30 +/-5 ℃, sample injection amount: 10 μ l, mobile phase with formic acid: the water system is a mobile phase A, acetonitrile is used as a mobile phase B, and gradient elution is adopted;

II: MS conditions: polarity: positive electrode, dry gas temperature: 320 ℃, dry air flow: 8L/min, atomizer pressure: 40psi, capillary voltage: 3500V, sheath gas temperature: 350 ℃, sheath gas flow rate: 10L/min, EMV: 200V, switching time setting of the switching valve:

0min：to Waste；

4.0min：to MS；

8.5min：to Waste。

2. a detection method of azidosartanbiphenyl is characterized in that: the detection method comprises the following steps:

(1) preparing solutions, namely respectively preparing a blank solution, a reference substance solution, a sensitivity solution and a test solution, wherein the reference substance solution comprises a sartanbiphenyl stock solution and a reference substance stock solution;

(2) the determination method comprises the following steps: measuring the content of the sartanbiphenyl azide by adopting LC-MS/MS, respectively adding a blank solution, a sensitivity solution, a reference solution and a test solution after the system is stable, and recording spectrograms;

i: the chromatographic conditions are as follows: octadecylsilane chemically bonded silica is used as a filling agent, the flow rate is 0.5mL/min +/-0.1 mL/min, the column temperature is as follows: 30 +/-5 ℃, sample injection amount: 10 μ l, mobile phase with formic acid: the water system is a mobile phase A, acetonitrile is used as a mobile phase B, and gradient elution is adopted;

II: MS conditions: polarity: positive electrode, dry gas temperature: 320 ℃, dry air flow: 8L/min, atomizer pressure: 40psi, capillary voltage: 3500V, sheath gas temperature: 350 ℃, sheath gas flow rate: 10L/min, EMV: 200V, switching time setting of the switching valve:

0min：to Waste；

4.0min：to MS；

8.5min：to Waste；

the blank solution is acetonitrile: water = 55: 45 (V/V);

the azidosartanbiphenyl stock solution: placing a proper amount of sartanbiphenyl azide reference substance into a volumetric flask, adding a proper amount of blank solution to dissolve and dilute the sartanbiphenyl azide reference substance to a scale, and shaking up;

the control stock solution: precisely measuring a proper amount of azidosartanbiphenyl stock solution, placing the azidosartanbiphenyl stock solution into a volumetric flask, adding a blank solution to dilute the azidosartanbiphenyl stock solution to a scale, and shaking up;

the control solution: measuring a proper amount of reference substance stock solution, placing the reference substance stock solution in a volumetric flask, adding the blank solution to dilute the reference substance stock solution to a scale, and shaking up;

the test solution is as follows: placing a proper amount of a sample in a volumetric flask, adding a blank solution to dissolve and dilute the sample to a scale, and shaking up;

the mobile phase is as follows:

mobile phase A: formic acid: water =1:1000 (V/V);

mobile phase B: acetonitrile;

the formic acid is HPLC grade;

the acetonitrile is of HPLC grade;

the water is ultrapure water;

the chromatographic column is Thermo Acclaim^TM RSLC120 C18，2.1×100mm，2.2 μm。

3. The method of claim 2, wherein the mobile phase gradient process is as follows:

。

4. a detection method of azido-sartanbiphenyl tetrazole comprises the following steps:

(1) preparing solutions, namely respectively preparing a diluent, a blank solution, a reference substance solution and a test solution, wherein the reference substance solution comprises an azide substituted sartanbiphenyl tetrazole stock solution and a reference substance stock solution;

(2) the determination method comprises the following steps: measuring the content of azido sartanbiphenyl tetrazole by adopting LC-MS/MS, after the system is stabilized, respectively adding a blank solution, a reference solution and a test solution, and recording spectrograms;

i: the chromatographic conditions are as follows: octadecylsilane chemically bonded silica is used as a filling agent, the flow rate is 0.5mL/min +/-0.1 mL/min, the column temperature is as follows: 30 +/-5 ℃, sample injection amount: 10 μ l, mobile phase with formic acid: the water system is a mobile phase A, acetonitrile is used as a mobile phase B, and gradient elution is adopted;

II: MS conditions: polarity: positive electrode, dry gas temperature: 320 ℃, dry air flow: 8L/min, atomizer pressure: 40psi, capillary voltage: 3500V, sheath gas temperature: 350 ℃, sheath gas flow rate: 10L/min, EMV: 200V, switching time setting of the switching valve:

0min：to Waste；

4.0min：to MS；

8.5min：to Waste；

the diluent is acetonitrile: water = 20: 80 (V/V);

the preparation steps of the blank solution are as follows: taking a proper amount of methanol in a volumetric flask, adding a diluent to dilute to a scale, and shaking up;

the nitrine sartanbiphenyl tetrazole stock solution comprises the following components: placing a proper amount of azide sartanbiphenyl tetrazole reference substance into a volumetric flask, adding a proper amount of methanol for dissolving and diluting to a scale, and shaking up;

the control stock solution: precisely measuring a proper amount of nitrine sartanbiphenyl tetrazole stock solution, placing the nitrine sartanbiphenyl tetrazole stock solution into a volumetric flask, adding methanol to dilute the solution to a scale, and shaking the solution uniformly;

the control solution: measuring a proper amount of reference substance stock solution, placing the reference substance stock solution in a volumetric flask, adding diluent to dilute the reference substance stock solution to a scale, and shaking up;

the test solution is as follows: placing a proper amount of a sample in a volumetric flask, adding methanol to dissolve and dilute the sample to a scale, and shaking up; precisely measuring a proper amount of the solution, placing the solution into a volumetric flask, adding a diluent to dilute the solution to a scale, and shaking up;

the mobile phase is as follows:

mobile phase A: formic acid: water =1:1000 (V/V);

mobile phase B: acetonitrile;

the formic acid is HPLC grade;

the methanol is HPLC grade;

the acetonitrile is of HPLC grade;

the water is ultrapure water;

the chromatographic column is Thermo Acclaim^TM RSLC120 C18，2.1×100mm，2.2 μm。

5. The method according to claim 4, wherein the mobile phase gradient process is as follows:

。

6. the method for detecting impurities in the synthesis of sartan drugs according to claim 1, wherein the method is used for detecting irbesartan, candesartan cilexetil or olmesartan medoxomil.

Technical Field

The invention belongs to the technical field of drug analysis, and particularly relates to a method for detecting impurities in sartan drug synthesis.

Background

During the synthesis of sartans, depending on the nature of the process of manufacture, by-products may be produced:wherein R is CN orAny one of the above.

Sartanbiphenyl azide, chemical name: 4'- (azidomethyl) - [1,1' -biphenyl ] -2-carbonitrile having the structural formula:

。

azido-sartanbiphenyl tetrazole, chemical name: 5- (4 '- (benzylazido) - [1,1' -biphenyl)]-2-yl) -1H-tetrazole of the formula:. During the synthesis of sartans, according to different production processes, sartansbiphenylene azide or sartansbiphenyltetrazole azide may be generated.

At present, few methods for detecting azides sartanbiphenyl or azides sartanbiphenyl tetrazole are reported, the method is very necessary to strictly control the content of impurities in products as a medical industry, the research on material impurities is beneficial to the optimization and quality control of the synthesis process of the drug, the method for detecting the azides sartanbiphenyl or azides sartanbiphenyl tetrazole in the sartanoid drug is researched, a better reference is provided for evaluating the quality of the sartanoid drug, a technical basis is also provided for optimizing the synthesis process of the sartanoid drug, and the method has great significance for determining adverse reactions caused by the azides sartanbiphenyl or azides sartanbiphenyl tetrazole in the sartanoid drug.

The invention provides a convenient, efficient and accurate detection method for solving the detection problem of azides sartanbiphenyl or azides sartanbiphenyl tetrazole, and the method can detect the content of the azides sartanbiphenyl or the azides sartanbiphenyl tetrazole in sartanas, so that the content of the azides sartanbiphenyl or the azides sartanbiphenyl tetrazole in the sartanas is effectively controlled, and the medication safety is ensured.

Disclosure of Invention

The invention provides a method for detecting azides sartanbiphenyl tetrazole, which is convenient, efficient and accurate, completely accords with the guide principle of Chinese pharmacopoeia method verification in the aspects of system applicability, repeatability, specificity and accuracy, and can be used for quality control of sartanas medicaments.

In order to achieve the purpose, the invention provides the following technical scheme:

a method for detecting impurities in the synthesis of sartan drugs, wherein the structural formula of the impurities is as follows:wherein R is CN orAny one of the above, the detection method comprising the steps of:

(1) preparing solutions, namely respectively preparing a diluent, a blank solution, a reference substance solution and a test solution, wherein the reference substance solution comprises an impurity stock solution and a reference substance stock solution.

(2) The determination method comprises the following steps: measuring the content of impurities by adopting LC-MS/MS, respectively adding a blank solution, a reference solution and a test solution after the system is stable, and recording spectrograms;

i: the chromatographic conditions are as follows: octadecylsilane chemically bonded silica is used as a filling agent, the flow rate is 0.5mL/min +/-0.1 mL/min, the column temperature is as follows: 30 +/-5 ℃, sample injection amount: 10 μ l, mobile phase with formic acid: the water system is a mobile phase A, acetonitrile is used as a mobile phase B, and gradient elution is adopted;

II: MS conditions: polarity: positive electrode, dry gas temperature: 320 ℃, dry air flow: 8L/min, atomizer pressure: 40psi, capillary voltage: 3500V, sheath gas temperature: 350 ℃, sheath gas flow rate: 10L/min, EMV: 200V, switching time setting of the switching valve:

0min：to Waste；

4.0min：to MS；

8.5min：to Waste。

a detection method of azidosartanbiphenyl is characterized in that: the detection method comprises the following steps:

(1) preparing solutions, namely respectively preparing a blank solution, a reference substance solution, a sensitivity solution and a test solution, wherein the reference substance solution comprises a sartanbiphenyl stock solution and a reference substance stock solution.

(2) The determination method comprises the following steps: measuring the content of the sartanbiphenyl azide by adopting LC-MS/MS, respectively adding a blank solution, a sensitivity solution, a reference solution and a test solution after the system is stable, and recording spectrograms;

i: the chromatographic conditions are as follows: octadecylsilane chemically bonded silica is used as a filling agent, the flow rate is 0.5mL/min +/-0.1 mL/min, the column temperature is as follows: 30 +/-5 ℃, sample injection amount: 10 μ l, mobile phase with formic acid: the water system is a mobile phase A, acetonitrile is used as a mobile phase B, and gradient elution is adopted;

II: MS conditions: polarity: positive electrode, dry gas temperature: 320 ℃, dry air flow: 8L/min, atomizer pressure: 40psi, capillary voltage: 3500V, sheath gas temperature: 350 ℃, sheath gas flow rate: 10L/min, EMV: 200V, switching time setting of the switching valve:

0min：to Waste；

4.0min：to MS；

8.5min：to Waste。

the blank solution is acetonitrile: water = 55: 45 (V/V);

the azidosartanbiphenyl stock solution: placing a proper amount of sartanbiphenyl azide reference substance into a volumetric flask, adding a proper amount of blank solution to dissolve and dilute the sartanbiphenyl azide reference substance to a scale, and shaking up;

the control stock solution: precisely measuring a proper amount of azidosartanbiphenyl stock solution, placing the azidosartanbiphenyl stock solution into a volumetric flask, adding a blank solution to dilute the azidosartanbiphenyl stock solution to a scale, and shaking up;

the control solution: measuring a proper amount of reference substance stock solution, placing the reference substance stock solution in a volumetric flask, adding the blank solution to dilute the reference substance stock solution to a scale, and shaking up;

the test solution is as follows: placing a proper amount of a sample in a volumetric flask, adding a blank solution to dissolve and dilute the sample to a scale, and shaking up;

the mobile phase is as follows:

mobile phase A: formic acid: water =1:1000 (V/V);

mobile phase B: acetonitrile;

the formic acid is HPLC grade;

the acetonitrile is of HPLC grade;

the water is ultrapure water;

the chromatographic column is Thermo Acclaim^TM RSLC120 C18，2.1×100mm，2.2 μm。

Further, the mobile phase gradient process is as follows:

。

a detection method of azido-sartanbiphenyl tetrazole comprises the following steps:

(1) preparing a solution, and respectively preparing a diluent, a blank solution, a reference solution and a test solution, wherein the reference solution comprises an azide oxatanbiphenyl tetrazole stock solution and a reference stock solution.

(2) The determination method comprises the following steps: measuring the content of azido sartanbiphenyl tetrazole by adopting LC-MS/MS, after the system is stabilized, respectively adding a blank solution, a reference solution and a test solution, and recording spectrograms;

i: the chromatographic conditions are as follows: octadecylsilane chemically bonded silica is used as a filling agent, the flow rate is 0.5mL/min +/-0.1 mL/min, the column temperature is as follows: 30 +/-5 ℃, sample injection amount: 10 μ l, mobile phase with formic acid: the water system is a mobile phase A, acetonitrile is used as a mobile phase B, and gradient elution is adopted;

II: MS conditions: polarity: positive electrode, dry gas temperature: 320 ℃, dry air flow: 8L/min, atomizer pressure: 40psi, capillary voltage: 3500V, sheath gas temperature: 350 ℃, sheath gas flow rate: 10L/min, EMV: 200V, switching time setting of the switching valve:

0min：to Waste；

4.0min：to MS；

8.5min：to Waste。

the diluent is acetonitrile: water = 20: 80 (V/V);

the preparation steps of the blank solution are as follows: taking a proper amount of methanol in a volumetric flask, adding a diluent to dilute to a scale, and shaking up;

the nitrine sartanbiphenyl tetrazole stock solution comprises the following components: placing a proper amount of azide sartanbiphenyl tetrazole reference substance into a volumetric flask, adding a proper amount of methanol for dissolving and diluting to a scale, and shaking up;

the control stock solution: precisely measuring a proper amount of nitrine sartanbiphenyl tetrazole stock solution, placing the nitrine sartanbiphenyl tetrazole stock solution into a volumetric flask, adding methanol to dilute the solution to a scale, and shaking the solution uniformly;

the control solution: measuring a proper amount of reference substance stock solution, placing the reference substance stock solution in a volumetric flask, adding diluent to dilute the reference substance stock solution to a scale, and shaking up;

the test solution is as follows: placing a proper amount of a sample in a volumetric flask, adding methanol to dissolve and dilute the sample to a scale, and shaking up; precisely measuring a proper amount of the solution, placing the solution into a volumetric flask, adding a diluent to dilute the solution to a scale, and shaking up;

the mobile phase is as follows:

mobile phase A: formic acid: water =1:1000 (V/V);

mobile phase B: acetonitrile;

the formic acid is HPLC grade;

the methanol is HPLC grade;

the acetonitrile is of HPLC grade;

the water is ultrapure water;

the chromatographic column is Thermo Acclaim^TM RSLC120 C18，2.1×100mm，2.2 μm。

Further, the mobile phase gradient process is as follows:

。

the method for detecting impurities in the synthesis of sartan drugs is used for detecting irbesartan, candesartan cilexetil or olmesartan medoxomil.

The invention provides a method for detecting impurities in the synthesis of sartan drugs for the first time, wherein the structural formula of the impurities is as follows:wherein R is CN orAny one of the two has the characteristics of high accuracy, high precision, good reproducibility, good stability, strong specificity and the like, and meanwhile, the invention adopts common reagents,the determination method has the advantages of simple operation, short time, low cost and short time consumption.

Drawings

FIG. 1 is a blank solution profile of example 2

FIG. 2 is a graph of the sensitivity solution of example 2

FIG. 3 is a spectrum of the control solution (a) of example 2

FIG. 4 is a solution profile of the control in example 2

FIG. 5 is a sample solution profile of example 3

FIG. 6 is the selective solution profile of example 3

FIG. 7 is LOD solution spectrum of example 4

Detailed Description

The invention is further illustrated by the following examples, which are not to be construed as limiting the invention thereto.

Example 1: detection of azides sartanbiphenyl tetrazole in olmesartan medoxomil

(1) Preparing a solution:

diluting liquid: acetonitrile: water = 20: 80 (V/V);

blank solution: and precisely measuring 500 mu l of methanol, putting the methanol into a 50ml measuring flask, adding a diluent to dilute the methanol to a scale, and shaking the methanol uniformly (the methanol can be prepared by increasing or reducing the volume according to the same proportion according to the actual use condition).

Nitrine sartanbiphenyl tetrazole stock solution: taking about 21mg of azide sartanbiphenyl tetrazole reference substance, precisely weighing, placing in a 100ml measuring flask, adding methanol for dissolving and diluting to scale, and shaking up; precisely measuring 500 mul of the solution, putting the solution into a 20ml measuring flask, adding methanol to dilute the solution to a scale, and shaking the solution uniformly; (concentration: 5.25 mug/ml)

Control stock solutions: accurately measuring 500 mu l of azide-sartanbiphenyl tetrazole stock solution, putting the nitrine-sartanbiphenyl tetrazole stock solution into a 50ml measuring flask, adding methanol to dilute the solution to a scale, and shaking the solution uniformly. (concentration: 52.5 ng/ml)

Control solution: and precisely measuring 500 mul of control stock solution, putting the control stock solution into a 50ml measuring flask, adding diluent to dilute the control stock solution to a scale, and shaking the control stock solution uniformly. (concentration: Azidosartan sartanbiphenyltetrazole 0.525 ng/ml)

Control solution (a): precisely measuring 1.0ml of the reference solution, placing the reference solution in a 10ml measuring flask, adding the blank solution to dilute to the scale, and shaking up. (concentration: Azide sartanbiphenyl tetrazole 0.0525 ng/ml)

Sensitivity solution: and precisely measuring 200 mul of the reference solution, putting the reference solution into a 10ml measuring flask, adding the blank solution to dilute to a scale, and shaking up. (concentration: azido sartanbiphenyl tetrazole 0.0105 ng/ml)

Test solution: taking about 70mg of a test sample, precisely weighing, placing in a 50ml measuring flask, adding a proper amount of methanol, ultrasonically dissolving, adding methanol to dilute to a scale, and shaking up; precisely measuring 500 mu l of the solution, putting the solution into a 50ml measuring flask, adding diluent to dilute the solution to a scale, and shaking the solution uniformly. (concentration: olmesartan medoxomil 0.014 mg/ml)

Selective solution: taking about 70mg of a sample to be tested, precisely weighing, placing the sample in a 50ml measuring flask, adding 500 microliters of azide substituted sartanbiphenyl tetrazole stock solution, adding a proper amount of methanol, ultrasonically dissolving, diluting the methanol to a scale, and shaking up; precisely measuring 500 mul of sample solution to be tested, putting the sample solution into a 50ml measuring flask, adding diluent to dilute the sample solution to a scale, and shaking up. (concentration: olmesartan medoxomil 0.014 mg/ml; azidate sartanbiphenyl tetrazole concentration: 0.525 ng/ml)

LOD solution: and precisely measuring 200 mul of the reference solution, putting the reference solution into a 10ml measuring flask, adding the blank solution to dilute to a scale, and shaking up. (concentration: azido sartanbiphenyl tetrazole 0.0105 ng/ml)

Remarking: the above solutions were all prepared at room temperature using a brown volumetric flask protected from light.

(2) The determination method comprises the following steps: after the system is stabilized, the blank solution 1 needle, the sensitivity solution 1 needle, the reference solution (a) 1 needle, the reference solution 6 needle, the blank solution 1 needle, the sample solution 1 needle are introduced, and the spectrogram is recorded.

Example 2: system applicability

The applicability of the system is realized by evaluating the signal-to-noise ratio (S/N) of the azides sartanbiphenyltetrazole in the sensitivity solution and the RSD of the peak area of the azides sartanbiphenyltetrazole in the 6-pin reference solution, and the S/N value of the azides sartanbiphenyltetrazole in the sensitivity solution is required to be not less than 3; RSD of the peak area of azidation sartanbiphenyl tetrazole in 6-needle reference solution is not more than 10.0%. In order to confirm the system applicability in the sequence running process, after the sample injection of the reference substance solution is completed, 1 needle of the reference substance solution is added every about 8 hours or at the end of the sequence, and the RSD of the azido sartanbiphenyl tetrazole in the reference substance solution of 6 continuous needles is required to be not more than 10.0 percent; if the range is exceeded, an evaluation survey should be conducted.

After the system was stabilized, the blank solution 1 needle, the sensitivity solution 1 needle, the control solution (a) 1 needle, and the control solution 6 needle were added and the spectra were recorded.

Example 3: specificity

The specificity is realized by measuring the separation degree between a nitrine sartanbiphenyl tetrazole peak and adjacent impurity peaks in a blank solution without interference on detection, and the recovery rate of the nitrine sartanbiphenyl tetrazole in the selective solution, wherein the blank solution is required to have no interference on detection, the separation degree between the nitrine sartanbiphenyl tetrazole peak and the adjacent impurity peaks in the selective solution is not less than 1.5, and the recovery rate of the nitrine sartanbiphenyl tetrazole in the selective solution is 80.0-120.0%.

After the system is stabilized, 1 needle of blank solution, test solution and reference solution is added, 3 needles of selective solution are added respectively, and the spectrogram is recorded.

Example 4: detection limit

The detection limit is determined by detecting that the ratio of the response signal to the noise is not less than 3: 1 to obtain. In the spectrum obtained, LOD is required_{Azide sartanbiphenyl tetrazole}Should not be more than 3.75ppm, and the S/N value of azido sartanbiphenyl tetrazole in the LOD solution should not be less than 3.

After the system is stabilized, 1 pin of blank solution and LOD solution are added respectively, and the spectrogram is recorded.

Example 5: durability

And observing the rule that the reference substance solution, the test solution and the selective solution are placed in the dark at room temperature for a period of time and then injected, wherein the change of the detection result along with the time provides reference for the placing time of the reference substance solution and the test solution during detection.

The method comprises the following steps: compared with the reference solution for 0hr, the recovery rate of the azides sartanbiphenyl tetrazole is between 80.0% and 120.0% during the room-temperature dark investigation of the reference solution, and no obvious change trend exists, so that the reference solution is stable during the room-temperature dark investigation;

if the sample solution is detected to be nitridized sartanbiphenyl tetrazole within 0hr, the sample solution is placed in a dark place at room temperature for a period of time, the change value of the measurement result is within 20% of the limit of the nitridized sartanbiphenyl tetrazole, and no obvious change trend exists, so that the sample solution is stable during the dark investigation at room temperature; if the sample solution is not detected with azides sartanbiphenyltetrazole within 0hr, and the sample solution is still not detected with azides sartanbiphenyltetrazole within a period of time of being placed at room temperature in the dark, the sample solution is stable during the room-temperature dark investigation;

the selective solution is placed at room temperature in a dark place for a period of time, the recovery rate of the azidation sartanbiphenyl tetrazole is between 80.0% and 120.0%, and no obvious change trend exists, so that the selective solution is stable during the room temperature in the dark place.

After the system is stabilized, after 0hr and at certain time intervals, the blank solution, the sample solution, the reference solution and the selective solution are respectively added for 1 needle, and the spectrogram is recorded.

Example 6: detection of azides sartanbiphenyltetrazole in candesartan cilexetil

(1) Solution preparation

Diluting liquid: acetonitrile: water = 20: 80 (V/V);

blank solution: and precisely measuring 500 mu l of methanol, putting the methanol into a 50ml measuring flask, adding a diluent to dilute the methanol to a scale, and shaking the methanol uniformly (the methanol can be prepared by increasing or reducing the volume according to the same proportion according to the actual use condition).

Nitrine sartanbiphenyl tetrazole stock solution: accurately weighing 47mg of azides sartanbiphenyl tetrazole reference substance, placing the reference substance into a 100ml measuring flask, adding methanol for dissolving, diluting to scale, and shaking up; precisely measuring 200 mul of the solution, putting the solution into a 20ml measuring flask, adding methanol to dilute the solution to a scale, and shaking the solution uniformly; (concentration: 4.7 mug/ml)

Control stock solutions: accurately measuring 500 mu l of azide-sartanbiphenyl tetrazole stock solution, putting the nitrine-sartanbiphenyl tetrazole stock solution into a 50ml measuring flask, adding methanol to dilute the solution to a scale, and shaking the solution uniformly. (concentration: 47 ng/ml)

Control solution: and precisely measuring 500 mul of control stock solution, putting the control stock solution into a 50ml measuring flask, adding diluent to dilute the control stock solution to a scale, and shaking the control stock solution uniformly. (concentration: Azidosartan sartanbiphenyltetrazole 0.47 ng/ml)

Control solution (a): precisely measuring 1.0ml of the reference solution, placing the reference solution in a 10ml measuring flask, adding the blank solution to dilute to the scale, and shaking up. (concentration: Azidosartan sartanbiphenyltetrazole 0.047 ng/ml)

Sensitivity solution: and precisely measuring 500 mul of the reference solution, putting the reference solution into a 20ml measuring flask, adding the blank solution to dilute to a scale, and shaking up. (concentration: 0.01175ng/ml oxatanbiphenyltetrazole)

Test solution: taking about 50mg of a test sample, precisely weighing, placing in a 50ml measuring flask, adding a proper amount of methanol, ultrasonically dissolving, adding methanol to dilute to a scale, and shaking up; precisely measuring 500 mu l of the solution, putting the solution into a 50ml measuring flask, adding diluent to dilute the solution to a scale, and shaking the solution uniformly. (concentration: candesartan cilexetil 0.01 mg/ml)

Remarking: the above solutions were all prepared at room temperature using a brown volumetric flask protected from light.

(2) The determination method comprises the following steps:

after the system is stabilized, the blank solution 1 needle, the sensitivity solution 1 needle, the reference solution (a) 1 needle, the reference solution 6 needle, the blank solution 1 needle, the sample solution 1 needle are introduced, and the spectrogram is recorded.

The detection of azides sartanbiphenyl tetrazole in candesartan cilexetil verifies the system adaptability, specificity, detection limit and durability, and the verification result is as follows:

example 7: detection of azidosartanbiphenyl in irbesartan

Blank solution: acetonitrile: water = 55: 45 (V/V);

azidosartanbiphenyl stock solutions: taking about 25mg of azidosartanbiphenyl reference, precisely weighing, placing in a 50ml measuring flask, adding a blank solution to dissolve and dilute to a scale, and shaking up. (concentration: 500 mug/ml)

Control stock solutions: precisely measuring 500 microliters of azidosartanbiphenyl stock solution, placing the azidosartanbiphenyl stock solution into a 20ml measuring flask, adding a blank solution to dilute the azidosartanbiphenyl stock solution to a scale, and shaking up; precisely measuring 1.0ml of the solution, placing the solution into a 100ml measuring flask, adding the blank solution to dilute to the scale, and shaking up. (concentration: 125 ng/ml)

Control solution: precisely measuring 1.0ml of the reference stock solution, placing the reference stock solution in a 50ml measuring flask, adding the blank solution to dilute to a scale, and shaking up. (concentration: Azidosartan biphenyl 2.5 ng/ml)

Sensitivity solution: precisely measuring 4.0ml of the reference solution, placing the reference solution in a 10ml measuring flask, adding the blank solution to dilute to the scale, and shaking up. (concentration: azidosartanbiphenyl 1 ng/ml)

Test solution: taking about 25mg of a sample, precisely weighing, placing in a 50ml measuring flask, adding a proper amount of blank solution, vortexing to dissolve and dilute the sample to a scale, and shaking up. (concentration: irbesartan 0.5 mg/ml)

After the system is stabilized, 1 needle of blank solution, 1 needle of sensitivity solution, 6 needles of reference solution, 1 needle of blank solution and 1 needle of test solution are added, and the spectrogram is recorded.

Results (ppm) = (RU/RS) × (CS/CU)

Wherein: RU: peak area of the sartanbiphenyl azide in the sample solution spectrum;

and RS: 6, the average peak area of the sartan azide and sartanbiphenyl in the solution spectrum of the reference substance;

CS: the concentration (ng/ml) of the azi-sartanbiphenyl in the control solution;

CU: irbesartan concentration (mg/ml) in test solution