CN115494184A - Method for simultaneously detecting methyl cumenesulfonate and ethyl cumenesulfonate in medicine - Google Patents
Method for simultaneously detecting methyl cumenesulfonate and ethyl cumenesulfonate in medicine Download PDFInfo
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Abstract
The invention provides a method for simultaneously detecting methyl cumenesulfonate and ethyl cumenesulfonate in a drug, which adopts an ultra-high performance liquid chromatography-mass spectrometer to respectively detect a reference solution and a test solution and calculates according to an external standard method to determine the contents of the methyl cumenesulfonate and the ethyl cumenesulfonate in a sample. The method adopts ultra-high performance liquid chromatography for separation, simultaneously detects the methyl cumenesulfonate and the ethyl cumenesulfonate by high-selectivity and high-sensitivity mass spectrum, has simple sample pretreatment, no interference of a sample matrix to the determination of a target peak, good durability and high detection efficiency.
Description
Technical Field
The invention relates to the field of drug analysis, in particular to a method for simultaneously detecting cumene methyl sulfonate and cumene ethyl sulfonate in a drug.
Background
Cumyl sulfonic acid or cumyl sulfonate is an important raw material in organic synthesis, which can form genotoxic impurities of methyl cumyl sulfonate and ethyl cumyl sulfonate with trace methanol and ethanol in the process, and the substances can generate alkylation reaction with DNA, thereby possibly becoming a cause for causing cancer, so that the control of the toxicological concern threshold (TTC) level of the impurities in the medicine is very important. The European Medical Evaluation Agency (EMEA), the U.S. Food and Drug Administration (FDA), and the international conference on drug registration (ICH) all impose limits on genotoxic impurities that must be controlled below the Threshold of Toxicological Concern (TTC) level. Wherein the structures of the isopropyl benzene sulfonic acid methyl ester and the isopropyl benzene sulfonic acid ethyl ester are as follows:
at present, no detection method for cumene sulfonate is found, and a method for simultaneously detecting cumene sulfonate and cumene sulfonate in a medicament is necessary to be developed.
Disclosure of Invention
The invention provides a method for simultaneously detecting methyl cumenesulfonate and ethyl cumenesulfonate in a medicament, which adopts ultra-high performance liquid chromatography-mass spectrometry to simultaneously detect the methyl cumenesulfonate and the ethyl cumenesulfonate, has high selectivity and high sensitivity, simple sample pretreatment, no interference of a sample matrix to the determination of a target peak and good durability, and comprises the following steps:
step 1: adding a diluent into a cumene sulfonic acid methyl ester and cumene sulfonic acid ethyl ester reference substance to prepare a reference substance solution;
step 2: adding a diluent into a sample to be tested to prepare a test solution;
and step 3: detecting the reference solution in the step 1 and the test solution in the step 2 by using an ultra-high performance liquid chromatography-mass spectrometer, and determining the contents of the cumene sulfonate and the cumene sulfonate ethyl in the sample according to an external standard method;
wherein, the chromatographic conditions of the ultra-high performance liquid chromatography-mass spectrometer in the step 3 are as follows:
a chromatographic column: octadecylsilane chemically bonded silica chromatographic column;
mobile phase: the mobile phase A is a volatile ammonium salt aqueous solution, preferably an ammonium formate aqueous solution or an ammonium acetate aqueous solution, and most preferably an ammonium formate aqueous solution; the concentration of the mobile phase A is 0.002-0.05 mol/L, preferably 0.005-0.05 mol/L, and most preferably 0.01mol/L;
the mobile phase B is methanol or acetonitrile, preferably methanol;
the elution mode of the mobile phase is gradient elution, and the procedure of the gradient elution is as follows:
| time (min) | Mobile phase A% | Mobile phase B% |
| 0.0 | 70 | 30 |
| 4.0 | 20 | 80 |
| 5.0 | 20 | 80 |
| 5.1 | 70 | 30 |
| 7.0 | 70 | 30 |
Preferably, the drug is selected from drugs using cumenesulfonic acid or cumenesulfonic acid salt in the process, and methanol or ethanol, preferably acarbose or micafungin sodium.
Preferably, the diluent used in step 1 or step 2 is methanol, acetonitrile, water or a mixture thereof, preferably an aqueous methanol solution or an aqueous acetonitrile solution, and most preferably an aqueous methanol solution.
Preferably, the diluent in step 1 or step 2 is 10% to 80% aqueous methanol or acetonitrile (V/V), preferably 10% to 60% aqueous methanol or acetonitrile (V/V), more preferably 20% to 50% aqueous methanol or acetonitrile (V/V), still more preferably 25% to 50% aqueous methanol or acetonitrile (V/V), and most preferably 30% aqueous methanol or acetonitrile (V/V).
Preferably, the diameter of the octadecylsilane chemically bonded silica chromatographic column in the step 3 is 2.1-4.6 mm, the length of the column is 50-100 mm, and the particle size is 1.6-3.5 μm; preferably, the diameter of the octadecylsilane chemically bonded silica chromatographic column is 2.1 mm-3.5 mm, the length of the column is 50 mm-100 mm, and the particle size is 1.6 mu m-2.5 mu m; more preferably, the diameter of the octadecylsilane chemically bonded silica chromatographic column is 2.1 mm-3.0 mm, the length of the column is 50 mm-100 mm, and the particle size is 1.7 μm-2.5 μm; most preferably, the octadecylsilane bonded silica gel chromatographic column is Agilent Eclipse Plus C 18 RRHD with a column diameter of 2.1mm and a column length of 50mm in sizeThe diameter is 1.8 μm.
Preferably, the column temperature of the chromatographic column in step 3 is from 25 ℃ to 45 ℃, preferably from 30 ℃ to 45 ℃, more preferably from 30 ℃ to 40 ℃, and most preferably 35 ℃.
Preferably, the flow rate of the mobile phase in step 3 is 0.2mL/min to 0.6mL/min, preferably 0.2mL/min to 0.5mL/min, more preferably 0.2mL/min to 0.4mL/min, and most preferably 0.3mL/min.
Preferably, the parameters of the mass spectrum in step 3 are as follows:
scanning mode: monitoring mass spectrum multiple reactions;
an ion source: an electrospray ion source;
ion source mode: a positive mode;
capillary voltage: 1.0 to 4.0KV, preferably 2.0 to 4.0KV, more preferably 3.5KV;
temperature of the drying gas: 300 to 650 ℃, preferably 350 to 600 ℃, more preferably 400 to 550 ℃, most preferably 500 ℃;
source temperature: at 150 ℃.
Preferably, the ultra-high performance liquid chromatography-mass spectrometer in the step 3 is a Waters ACQUITY I UPLC Class-TQ-S micro or Agilent 1290-6470.
Compared with the prior art, the invention has the following beneficial effects:
the content of the cumene sulfonic acid methyl ester and the cumene sulfonic acid ethyl ester in the sample can be detected simultaneously; the sample pretreatment is simple, and the sample adopts a direct dissolution mode without derivatization treatment; the separation degree is good, and the cumene sulfonic acid methyl ester and the cumene sulfonic acid ethyl ester are completely and effectively separated from the sample in 7 min; the linear correlation coefficient is good, and the linear correlation coefficients r of the cumene sulfonic acid methyl ester and the cumene sulfonic acid ethyl ester are both more than or equal to 0.999 within the respective linear range; the sensitivity is high, the lowest detection limit of a cumene sulfonate instrument is 5.7350ng/mL, the lowest detection limit of an cumene sulfonate instrument is 4.9500ng/mL, the lowest detection limit of cumene sulfonate is 1.7205ng/mL, and the lowest detection limit of cumene sulfonate is 1.4850ng/mL; the recovery rate is high, and the standard addition accuracy recovery rates of the isopropyl benzene methyl sulfonate and the isopropyl benzene ethyl sulfonate are both 90-115%. The method can be used for simultaneously detecting two kinds of cumenesulfonates, so that the detection efficiency is improved, meanwhile, the method adopts high-selectivity and high-sensitivity mass spectrum for detection, the pretreatment of the sample is simple, and the sample matrix has no interference on the determination of a target peak; the method has good durability and high detection efficiency, and can well detect the contents of the cumene sulfonate and the cumene sulfonate ethyl ester in the sample so as to ensure the product quality.
Drawings
FIG. 1 is a super-performance liquid chromatography-mass spectrometry combined spectrum of blank solvents of methyl cumenesulfonate and ethyl cumenesulfonate.
FIG. 2 is a diagram of a reference solution of cumene sulfonate and ethyl cumenesulfonate in ultra-high performance liquid chromatography-mass spectrometry.
FIG. 3 is a linear spectrum of methyl cumenesulfonate according to the present invention.
FIG. 4 is a linear spectrum of ethyl cumenesulfonate according to the present invention.
Detailed Description
The following examples are intended to further illustrate the present invention, but it should be understood that the following examples are only illustrative of the present invention and are not to be construed as limiting the scope of the present invention, which is defined by the appended claims.
In the following examples, all temperatures are in degrees celsius unless otherwise indicated; unless otherwise indicated, various starting materials and reagents were obtained commercially and were used without further purification; unless otherwise indicated, each solvent is a technical grade solvent and is used without further treatment. The methanol is commercially available chromatographic purity, the ammonium formate is commercially available mass spectrometric purity, the water is self-made ultrapure water, and the purity of the isopropyl benzene methyl sulfonate and the purity of the isopropyl benzene ethyl sulfonate reference products are both 98% (both purchased from Shanghai Limited liability company of Sandy Yam medicine technology).
Preparation of test solution
Respectively weighing a proper amount of acarbose or micafungin sodium samples, dissolving and diluting the samples with 30% methanol aqueous solution (V/V) to obtain a solution containing 4mg/mL acarbose or micafungin sodium, and taking the solution as a test solution.
Preparation of control stock solutions
Accurately weighing appropriate reference substances of the isopropylbenzene methyl sulfonate and the isopropylbenzene ethyl sulfonate respectively, and dissolving and diluting the reference substances by using a diluent to obtain reference substance stock solutions with the concentration of both the isopropylbenzene methyl sulfonate and the isopropylbenzene ethyl sulfonate being 50 ng/mL.
Example 1
Detection condition of ultra-high performance liquid phase-mass spectrometry
The instrument comprises the following steps: waters ACQUITY I UPLC Class-TQ-Smicro)
Chromatographic conditions are as follows:
a chromatographic column: agilent Eclipse Plus C 18 RRHD 50*2.1mm 1.8μm
Flow rate: 0.3mL/min
Column temperature: 35 deg.C
Sample introduction amount: 5.0. Mu.L
Mobile phase A:0.01mol/L ammonium formate aqueous solution
Mobile phase B: methanol
Diluent agent: 30% aqueous methanol solution (V/V)
Blank solvent: diluting agent
Gradient:
mass spectrum conditions:
(1) Specificity experiments
a) Blank solvent: 30% methanol aqueous solution (V/V), the characteristic ion diagram of which is shown in FIG. 1;
b) Control solution: transferring 4.0mL of the reference stock solution into a 10mL volumetric flask, diluting the reference stock solution to a scale with a diluent, and uniformly mixing, wherein the characteristic ion diagram is shown in FIG. 2;
c) Adding a standard test solution: weighing 40mg of acarbose sample into a 10mL volumetric flask, adding 4.0mL of reference stock solution, dissolving with a proper amount of diluent, diluting with the diluent to scale, and mixing uniformly.
Weighing a micafungin sodium sample 40mg in a 10mL volumetric flask, adding 4.0mL of a reference substance stock solution, dissolving with a proper amount of diluent, diluting with the diluent to a scale, and mixing uniformly.
The detection method is characterized in that a blank solvent, a reference substance solution and a special solution of a sample solution added with a standard are detected according to the method, the detection results are shown in table 1, the blank solvent and the sample solution have no interference on the detection of the methyl cumenesulfonate and the ethyl cumenesulfonate, and the special property of the ultra-high performance liquid-mass spectrometry detection method is good.
Table 1 example 1 results of the determination of the specialty solutions
(2) Detection limit and quantification limit
a) Quantitative limiting solution: transferring 1.0mL of the reference substance stock solution into 6 10mL volumetric flasks, diluting the reference substance stock solution to a scale with a diluent, and uniformly mixing;
b) Detection limiting solution: transferring 3.0mL of the first quantitative limiting solution into a 10mL volumetric flask, diluting the solution to a scale with a diluent, and uniformly mixing;
the quantitative limiting solution and the detection limiting solution are detected according to the method, the signal-to-noise ratio (S/N) of the quantitative limiting solution is required to be more than or equal to 10, the detection limiting solution (S/N) is required to be more than or equal to 3, and the detection results are shown in tables 2 and 3:
TABLE 2 EXAMPLE 1 ultra high performance liquid Mass Spectrometry quantitation Limit
TABLE 3 example 1 ultra performance liquid chromatography-mass spectrometry combined detection limit
(3) Linearity
Linear solution: transferring 1.0mL, 2.0mL, 4.0mL, 6.0mL and 8.0mL of cumene sulfonate stock solutions, respectively placing the transfer solutions in 5 10mL volumetric flasks, diluting the transfer solutions to a scale with a diluent, uniformly mixing, and preparing two parts in parallel for each linear concentration.
One injection of each linear solution was performed according to the above method, and the linear equation and the linear correlation coefficient were calculated, and the results are shown in table 4.
TABLE 4 example 1 detection method of ultra performance liquid chromatography-mass spectrometer with linear regression equation and linear correlation coefficient r
According to the experimental results, the high performance liquid chromatography-mass spectrometer detection method has good linear correlation in the detection range, the linear correlation coefficients r are all 0.9999, and the contents of the cumene sulfonate methyl and the cumene sulfonate ethyl can be accurately quantified.
(4) Accuracy and repeatability experiments
a) Experiment of accuracy
Preparing acarbose or micafungin sodium test solution, respectively adding three reference solutions of cumene sulfonate and cumene sulfonate ethyl ester with different concentration levels, and preparing three parts at each concentration in parallel, wherein the preparation concentrations are shown in tables 5 and 6. Each portion of the spiked sample solution was analyzed by sampling as described above, and the recovery rate of each portion of the solution and the average recovery rate of each concentration level were calculated, and the results are shown in tables 5 and 6.
TABLE 5 example 1 detection method accuracy of ultra performance liquid chromatography-mass spectrometer for micafungin sodium
TABLE 6 example 1 detection method accuracy of ultra performance liquid chromatography-mass spectrometer for acarbose
b) Repeatability test
6 parts of solutions with moderate concentration levels of standard acarbose or standard micafungin sodium are prepared, each part of solution is subjected to sample injection analysis once according to the method, and the RSD of the recovery rate of the 6 parts of standard solutions is calculated. The results are shown in tables 7 and 8.
TABLE 7 example 1 detection method repeatability of ultra performance liquid chromatography-mass spectrometer in micafungin sodium
TABLE 8 example 1 detection method repeatability of ultra performance liquid chromatography-mass spectrometer for acarbose
According to the experimental results, the detection method of the high performance liquid chromatography-mass spectrometer has the advantages that the recovery rate of the high performance liquid chromatography-mass spectrometer is 90-115% in the accuracy and repeatability experiment, the RSD is less than 5%, and the accuracy and the repeatability are good.
(5) Detection of samples of methyl cumenesulfonate and ethyl cumenesulfonate in acarbose or micafungin sodium
Weighing 40mg of acarbose sample in a 10mL volumetric flask, dissolving the acarbose sample by using a proper amount of diluent, then diluting the acarbose sample to a scale by using the diluent, and uniformly mixing; weighing a micafungin sodium sample 40mg in a 10mL volumetric flask, dissolving the micafungin sodium sample with a proper amount of diluent, then diluting the sample with the diluent to a scale, and uniformly mixing the sample, wherein detection results are shown in tables 9 and 10.
TABLE 9 EXAMPLE 1 test results of micafungin sodium sample
TABLE 10 example 1 acarbose sample test results
From the above experimental results, it is found that methyl cumenesulfonate and ethyl cumenesulfonate were not detected in both acarbose and micafungin samples, and both were reported to be less than the limit of detection.
Example 2
Detection condition of ultra-high performance liquid phase-mass spectrometry
The instrument comprises the following steps: agilent 1290-6470
Chromatographic conditions are as follows:
a chromatographic column: agilent Eclipse Plus C 18 RRHD 50*2.1mm 1.8μm
Flow rate: 0.3mL/min
Column temperature: 35 deg.C
Sample injection amount: 5.0. Mu.L
Mobile phase A:0.01moL/L ammonium formate aqueous solution
Mobile phase B: methanol
Diluent agent: 30% aqueous methanol solution (V/V)
Blank solvent: diluting agent
Gradient:
| time (min) | Mobile phase A% | Mobile phase B% |
| 0.0 | 70 | 30 |
| 4.0 | 20 | 80 |
| 5.0 | 20 | 80 |
| 5.1 | 70 | 30 |
| 7.0 | 70 | 30 |
Mass spectrum conditions:
(1) Specificity experiments
a) Blank solvent: 30% aqueous methanol (V/V);
b) Control solution: transferring 4.0mL of the reference substance stock solution into a 10mL volumetric flask, diluting the reference substance stock solution to a scale with a diluent, and uniformly mixing;
c) Adding a standard test solution: weighing 40mg of acarbose sample into a 10mL volumetric flask, adding 4.0mL of reference stock solution, dissolving with a proper amount of diluent, diluting with the diluent to scale, and mixing uniformly.
Weighing 40mg of micafungin sodium sample into a 10mL volumetric flask, adding 4.0mL of reference substance stock solution, dissolving with a proper amount of diluent, diluting with the diluent to scale, and mixing uniformly.
The detection results are shown in table 11, and the experimental results show that the blank solvent and the sample solution do not interfere with the detection of the methyl cumenesulfonate and the ethyl cumenesulfonate, and the specificity of the ultra-performance liquid-mass spectrometry detection method is good.
Table 11 example 2 results of the specific solution assay
| Item | Cumene sulfonic acid methaneEsters | Isopropyl benzene sulfonic acid ethyl ester |
| Retention time in blank solvent (min) | ND | ND |
| Control solution Retention time (min) | 2.72 | 3.95 |
| Retention time in spiked solution (min) | 2.72 | 3.95 |
| Retention time in sample solution (min) | ND | ND |
(2) Detection limit and quantification limit
a) Quantitative limiting solution: transferring 1.0mL of the reference substance stock solution into 6 10mL volumetric flasks, diluting the reference substance stock solution to a scale with a diluent, and uniformly mixing;
b) Detection limiting solution: transferring 3.0mL of the first quantitative limiting solution into a 10mL volumetric flask, diluting the solution to a scale with a diluent, and uniformly mixing;
the quantitative limiting solution and the detection limiting solution are detected according to the method, the signal-to-noise ratio (S/N) of the quantitative limiting solution is required to be more than or equal to 10, the detection limiting solution (S/N) is required to be more than or equal to 3, and the detection results are shown in tables 12 and 13:
TABLE 12 example 2 ultra performance liquid chromatography-mass spectrometry combined quantitation limit
TABLE 13 EXAMPLE 2 ultra performance liquid Mass Spectrometry detection limits
(3) Linearity
Linear solution: transferring 1.0mL, 2.0mL, 4.0mL, 6.0mL and 8.0mL of cumene sulfonate stock solutions, respectively placing the transfer solutions in 5 10mL volumetric flasks, diluting the transfer solutions to a scale with a diluent, uniformly mixing, and preparing two parts in parallel for each linear concentration.
One injection of each linear solution was performed according to the above method, and the linear equation and the linear correlation coefficient were calculated, and the results are shown in table 14.
TABLE 14 example 2 detection method of ultra performance liquid chromatography-mass spectrometer with linear regression equation and linear correlation coefficient r
From the experimental results, the high performance liquid chromatography-mass spectrometer detection method has good linear relation in the detection range, the linear correlation coefficients are 0.9998 and 0.9999, and the contents of the methyl cumenesulfonate and the ethyl cumenesulfonate can be accurately quantified.
(4) Accuracy and repeatability experiments
a) Experiment of accuracy
Preparing acarbose or micafungin sodium test solution, adding three reference solutions of cumene sulfonate and cumene sulfonate ethyl ester with different concentration levels, and preparing three parts of each concentration in parallel, wherein the preparation concentrations are shown in tables 15 and 16. Each portion of the sample solution was analyzed by the above method, and the recovery rate of each portion of the sample solution and the average recovery rate of each concentration level were calculated. The results are shown in tables 15 and 16.
TABLE 15 example 2 detection method accuracy of ultra performance liquid chromatography-mass spectrometer for micafungin sodium
TABLE 16 example 2 detection method accuracy of ultra performance liquid chromatography-mass spectrometer for acarbose
b) Repeatability test
6 parts of solutions with moderate concentration levels of standard acarbose or standard micafungin sodium are prepared, each part of solution is subjected to sample injection analysis once according to the method, and the RSD of the recovery rate of the 6 parts of standard solutions is calculated. The results are shown in tables 17 and 18.
TABLE 17 example 2 detection method repeatability of ultra performance liquid chromatography-mass spectrometer in micafungin sodium
TABLE 18 example 2 detection method repeatability of ultra performance liquid chromatography-mass spectrometer for acarbose
According to the experimental results, the detection method of the high performance liquid chromatography-mass spectrometer has the advantages that the accuracy and the repeatability experiment recovery rate are 95% -105%, the RSD is less than 5%, and the accuracy and the repeatability are good.
(5) Detection of samples of methyl cumenesulfonate and ethyl cumenesulfonate in acarbose or micafungin sodium
Weighing 40mg of acarbose sample in a 10mL volumetric flask, dissolving the acarbose sample by using a proper amount of diluent, then diluting the acarbose sample to a scale by using the diluent, and uniformly mixing; weighing a micafungin sodium sample 40mg in a 10mL volumetric flask, dissolving the micafungin sodium sample with a proper amount of diluent, then diluting the sample with the diluent to a scale, and uniformly mixing the sample, wherein detection results are shown in tables 19 and 20.
TABLE 19 example 2 Micafungin sodium sample test results
TABLE 20 example 2 acarbose sample test results
From the above experimental results, it is found that methyl cumenesulfonate and ethyl cumenesulfonate were not detected in both acarbose and micafungin samples, and both were reported to be less than the limit of detection.
Comparative example 1 Mobile phase A was 0.1% (V/V) aqueous formic acid and Mobile phase B was methanol
0.01moL/L of the aqueous ammonium formate solution of example 1 was replaced by 0.1% (V/V) aqueous formic acid solution, and the other chromatographic mass spectrometry conditions (example 1) were unchanged. 3 parts of medium-concentration standard acarbose solution are prepared, each part of solution is subjected to sample injection and analysis once, a chromatogram is recorded, and the result is shown in Table 21. The results show that when 0.1% (V/V) formic acid aqueous solution is used as the mobile phase, the recovery rate of the sample standard addition increases with time, and the recovery rate is lower and lower, and exceeds the recovery rate requirement of a laboratory (the required recovery rate is 50-150%).
TABLE 21 recovery of spiked solutions of acarbose medium concentration with 0.1% (V/V) formic acid in water as mobile phase
Comparative example 2 the column length of the column was 150mm
The chromatographic column was replaced with Waters ACQUITY UPLC BEH C 18 150mm x 2.1mm 1.8 μm, other chromatographic mass spectrometry conditions (example 1) were not changed, and methyl cumyl sulfonate and ethyl cumyl sulfonate control solutions were injected, and no target peaks were found, indicating that the target peaks of methyl cumyl sulfonate and ethyl cumyl sulfonate remained strong and did not elute.
Claims (9)
1. A method for simultaneously detecting methyl cumenesulfonate and ethyl cumenesulfonate in a drug is characterized by comprising the following steps:
step 1: adding a diluent into a reference product of the cumene sulfonic acid methyl ester and the cumene sulfonic acid ethyl ester to prepare a reference product solution;
step 2: adding a diluent into a sample to be tested to prepare a test solution;
and step 3: detecting the reference solution in the step 1 and the test solution in the step 2 by using an ultra-high performance liquid chromatography-mass spectrometer, and determining the contents of the cumene sulfonate and the cumene sulfonate ethyl in the sample according to an external standard method;
wherein, the chromatographic conditions of the ultra-high performance liquid chromatography-mass spectrometer in the step 3 are as follows:
and (3) chromatographic column: octadecylsilane chemically bonded silica chromatographic column;
mobile phase: the mobile phase A is a volatile ammonium salt aqueous solution, preferably an ammonium formate aqueous solution or an ammonium acetate aqueous solution, and most preferably an ammonium formate aqueous solution; the concentration of the mobile phase A is 0.002-0.05 mol/L, preferably 0.005-0.05 mol/L, and most preferably 0.01mol/L;
the mobile phase B is methanol or acetonitrile, preferably methanol;
the elution mode of the mobile phase is gradient elution, and the procedure of the gradient elution is as follows:
2. The method according to claim 1, wherein the drug is selected from the group consisting of cumenesulfonic acid or cumenesulfonic acid salt used in the process, and a drug using methanol or ethanol, preferably acarbose or micafungin sodium.
3. A process according to claim 1 or 2, characterized in that the diluent in step 1 or step 2 is methanol, acetonitrile, water or a mixture thereof, preferably aqueous methanol or acetonitrile, most preferably aqueous methanol.
4. A process according to any one of claims 1 to 3, wherein the diluent in step 1 or step 2 is 10% to 80% aqueous methanol or acetonitrile (V/V), preferably 10% to 60% aqueous methanol or acetonitrile (V/V), more preferably 20% to 50% aqueous methanol or acetonitrile (V/V), even more preferably 25% to 50% aqueous methanol or acetonitrile (V/V), and most preferably 30% aqueous methanol or acetonitrile (V/V).
5. The method according to any one of claims 1 to 4, wherein the octadecylsilane-bonded silica chromatographic column in step 3 has a column diameter of 2.1mm to 4.6mm, a column length of 50mm to 100mm, and a particle diameter of 1.6 μm to 3.5 μm; preferably, the diameter of the octadecylsilane chemically bonded silica chromatographic column is 2.1 mm-3.5 mm, the length of the column is 50 mm-100 mm, and the particle size is 1.6 mu m-2.5 mu m; more preferably, the octadecylsilane chemically bonded silica chromatographic column has a column diameter of 2.1mm to 3.0mm, a column length of 50mm to 100mm, and a particle diameter of 1.7 μm to 2.5 μm; most preferably, the octadecylsilane bonded silica chromatographic column is Agilent Eclipse Plus C 18 RRHD, column diameter 2.1mm, column length 50mm, particle size 1.8 μm.
6. A method according to any one of claims 1 to 5, wherein the column temperature of the chromatography column in step 3 is from 25 ℃ to 45 ℃, preferably from 30 ℃ to 45 ℃, more preferably from 30 ℃ to 40 ℃, most preferably 35 ℃.
7. The method according to any one of claims 1 to 6, wherein the flow rate of the mobile phase in step 3 is from 0.2mL/min to 0.6mL/min, preferably from 0.2mL/min to 0.5mL/min, more preferably from 0.2mL/min to 0.4mL/min, most preferably 0.3mL/min.
8. The method according to any one of claims 1 to 7, wherein the parameters of the mass spectrum in step 3 are:
scanning mode: monitoring mass spectrum multiple reactions;
an ion source: an electrospray ion source;
ion source mode: a positive mode;
capillary voltage: 1.0 to 4.0KV, preferably 2.0 to 4.0KV, more preferably 3.5KV;
temperature of the drying gas: 300 to 650 ℃, preferably 350 to 600 ℃, more preferably 400 to 550 ℃, most preferably 500 ℃;
source temperature: at 150 ℃.
9. The method according to any one of claims 1 to 8, wherein the hplc-ms in step 3 is Waters acquisition I UPLC Class-TQ-S micro or Agilent 1290-6470.
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Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1521176A (en) * | 2003-02-13 | 2004-08-18 | 浙江海正药业股份有限公司 | Method for producing highly pure acarbose |
| US20040266644A1 (en) * | 2002-03-15 | 2004-12-30 | Michael Seebach | Ammonium nitriles and the use thereof as hydrophobic bleaching activators |
| US20080027083A1 (en) * | 2006-07-28 | 2008-01-31 | Bristol-Myers Squibb Company | Modulators of chemokine receptor activity crystalline forms and process |
| US20140114049A1 (en) * | 2011-05-12 | 2014-04-24 | Shanghai Techwell Biopharmaceutical Co., Ltd. | Preparation method of micafungin sodium |
| US20150065417A1 (en) * | 2012-03-30 | 2015-03-05 | Shanghai Techwell Biophamaceutical Co., Ltd. | High purity cyclopeptide compound as well as preparation method and use thereof |
| CN106336349A (en) * | 2016-08-24 | 2017-01-18 | 南开大学 | Bromotrifluoromethoxy compound and synthetic method thereof |
| US20170313792A1 (en) * | 2014-10-28 | 2017-11-02 | The University Of Tokyo | Method for producing polar group-containing olefin polymer |
| CN111122724A (en) * | 2019-12-17 | 2020-05-08 | 纳谱分析技术（苏州）有限公司 | Method for analyzing acarbose and related substances |
| CN112903838A (en) * | 2021-01-15 | 2021-06-04 | 浙江海正药业股份有限公司 | Method for determining related substances in Favilavir |
| CN114113346A (en) * | 2020-08-25 | 2022-03-01 | 石药集团恩必普药业有限公司 | Detection method for ethyl p-toluenesulfonate and isopropyl p-toluenesulfonate in dabigatran etexilate bulk drug or preparation |
-
2022
- 2022-05-09 CN CN202210506801.XA patent/CN115494184B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040266644A1 (en) * | 2002-03-15 | 2004-12-30 | Michael Seebach | Ammonium nitriles and the use thereof as hydrophobic bleaching activators |
| CN1521176A (en) * | 2003-02-13 | 2004-08-18 | 浙江海正药业股份有限公司 | Method for producing highly pure acarbose |
| US20080027083A1 (en) * | 2006-07-28 | 2008-01-31 | Bristol-Myers Squibb Company | Modulators of chemokine receptor activity crystalline forms and process |
| US20140114049A1 (en) * | 2011-05-12 | 2014-04-24 | Shanghai Techwell Biopharmaceutical Co., Ltd. | Preparation method of micafungin sodium |
| US20150065417A1 (en) * | 2012-03-30 | 2015-03-05 | Shanghai Techwell Biophamaceutical Co., Ltd. | High purity cyclopeptide compound as well as preparation method and use thereof |
| US20170313792A1 (en) * | 2014-10-28 | 2017-11-02 | The University Of Tokyo | Method for producing polar group-containing olefin polymer |
| CN106336349A (en) * | 2016-08-24 | 2017-01-18 | 南开大学 | Bromotrifluoromethoxy compound and synthetic method thereof |
| CN111122724A (en) * | 2019-12-17 | 2020-05-08 | 纳谱分析技术（苏州）有限公司 | Method for analyzing acarbose and related substances |
| CN114113346A (en) * | 2020-08-25 | 2022-03-01 | 石药集团恩必普药业有限公司 | Detection method for ethyl p-toluenesulfonate and isopropyl p-toluenesulfonate in dabigatran etexilate bulk drug or preparation |
| CN112903838A (en) * | 2021-01-15 | 2021-06-04 | 浙江海正药业股份有限公司 | Method for determining related substances in Favilavir |
Non-Patent Citations (5)
| Title |
|---|
| DAVID J. SNODIN: "Elusive Impurities-Evidence versus Hypothesis. Technical and Regulatory Update on Alkyl Sulfonates in Sulfonic Acid Salts", ORGANIC PROCESS RESEARCH & DEVELOPMENT, vol. 23 * |
| 和燕玲;邵伍军;金美春;郑晓鹤;: "HP LC-QTOF法测定米卡芬净钠中EDC・HCl的含量", 中兽医医药杂志, no. 05 * |
| 张霁;张英俊;聂飚;: "药物研发中基因毒性杂质的控制策略与方法探索进展", 中国医药工业杂志, no. 09 * |
| 彭远松,魏承志: "HPLC测定阿卡波糖胶囊的含量及有关物质", 华西药学杂志, no. 01 * |
| 武晗燕;周长明;段艳;姚瑛;: "阿卡波糖有关物质的方法学研究", 生命科学仪器, no. 06 * |
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