CN112034056A - Detection method for detecting tetrabutylammonium bromide content in levetiracetam - Google Patents
Detection method for detecting tetrabutylammonium bromide content in levetiracetam Download PDFInfo
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- JRMUNVKIHCOMHV-UHFFFAOYSA-M tetrabutylammonium bromide Chemical compound [Br-].CCCC[N+](CCCC)(CCCC)CCCC JRMUNVKIHCOMHV-UHFFFAOYSA-M 0.000 title claims abstract description 47
- 238000001514 detection method Methods 0.000 title claims abstract description 24
- 229960004002 levetiracetam Drugs 0.000 title claims abstract description 15
- HPHUVLMMVZITSG-ZCFIWIBFSA-N levetiracetam Chemical compound CC[C@H](C(N)=O)N1CCCC1=O HPHUVLMMVZITSG-ZCFIWIBFSA-N 0.000 title claims abstract description 15
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 99
- 239000003814 drug Substances 0.000 claims abstract description 38
- 238000000034 method Methods 0.000 claims abstract description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 13
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 11
- 239000012071 phase Substances 0.000 claims abstract description 11
- 150000003839 salts Chemical class 0.000 claims abstract description 10
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 6
- 238000010811 Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry Methods 0.000 claims abstract description 3
- 239000000945 filler Substances 0.000 claims abstract description 3
- 238000004949 mass spectrometry Methods 0.000 claims abstract description 3
- 238000002156 mixing Methods 0.000 claims abstract description 3
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims abstract description 3
- 239000012074 organic phase Substances 0.000 claims abstract description 3
- 238000005173 quadrupole mass spectroscopy Methods 0.000 claims abstract description 3
- 239000000377 silicon dioxide Substances 0.000 claims abstract description 3
- 238000001195 ultra high performance liquid chromatography Methods 0.000 claims abstract description 3
- 239000000243 solution Substances 0.000 claims description 33
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 21
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 16
- 238000007865 diluting Methods 0.000 claims description 14
- 150000002500 ions Chemical class 0.000 claims description 14
- 229940079593 drug Drugs 0.000 claims description 13
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 8
- 235000019253 formic acid Nutrition 0.000 claims description 8
- 238000001819 mass spectrum Methods 0.000 claims description 8
- 239000013558 reference substance Substances 0.000 claims description 7
- 239000000706 filtrate Substances 0.000 claims description 6
- 239000012085 test solution Substances 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 5
- 238000006243 chemical reaction Methods 0.000 claims description 4
- 239000012528 membrane Substances 0.000 claims description 4
- VZTDIZULWFCMLS-UHFFFAOYSA-N ammonium formate Chemical group [NH4+].[O-]C=O VZTDIZULWFCMLS-UHFFFAOYSA-N 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 238000012544 monitoring process Methods 0.000 claims description 3
- 239000012088 reference solution Substances 0.000 claims description 3
- 239000001961 anticonvulsive agent Substances 0.000 claims description 2
- 238000010828 elution Methods 0.000 claims description 2
- 238000010812 external standard method Methods 0.000 claims description 2
- 239000012634 fragment Substances 0.000 claims description 2
- 239000007921 spray Substances 0.000 claims description 2
- 238000005406 washing Methods 0.000 claims description 2
- 229960003965 antiepileptics Drugs 0.000 claims 1
- 239000008346 aqueous phase Substances 0.000 claims 1
- 230000035945 sensitivity Effects 0.000 abstract description 5
- 238000002360 preparation method Methods 0.000 abstract description 3
- 238000003908 quality control method Methods 0.000 abstract description 2
- 239000011550 stock solution Substances 0.000 description 18
- 239000012535 impurity Substances 0.000 description 13
- 239000000523 sample Substances 0.000 description 13
- 230000001738 genotoxic effect Effects 0.000 description 8
- 238000004090 dissolution Methods 0.000 description 7
- 231100000024 genotoxic Toxicity 0.000 description 7
- CPELXLSAUQHCOX-UHFFFAOYSA-M Bromide Chemical compound [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 4
- 229940006460 bromide ion Drugs 0.000 description 4
- 238000004255 ion exchange chromatography Methods 0.000 description 4
- 239000002904 solvent Substances 0.000 description 3
- AHOUBRCZNHFOSL-UHFFFAOYSA-N 3-(1,3-benzodioxol-5-yloxymethyl)-4-(4-fluorophenyl)piperidine Chemical compound C1=CC(F)=CC=C1C1C(COC=2C=C3OCOC3=CC=2)CNCC1 AHOUBRCZNHFOSL-UHFFFAOYSA-N 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 238000012417 linear regression Methods 0.000 description 2
- 238000004811 liquid chromatography Methods 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- JVHXJTBJCFBINQ-ADAARDCZSA-N Dapagliflozin Chemical compound C1=CC(OCC)=CC=C1CC1=CC([C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)=CC=C1Cl JVHXJTBJCFBINQ-ADAARDCZSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- 230000003178 anti-diabetic effect Effects 0.000 description 1
- 230000003556 anti-epileptic effect Effects 0.000 description 1
- 230000002924 anti-infective effect Effects 0.000 description 1
- 239000003472 antidiabetic agent Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 108091092356 cellular DNA Proteins 0.000 description 1
- 229960003834 dapagliflozin Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 231100000025 genetic toxicology Toxicity 0.000 description 1
- 238000002013 hydrophilic interaction chromatography Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 238000010829 isocratic elution Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 231100000219 mutagenic Toxicity 0.000 description 1
- 230000003505 mutagenic effect Effects 0.000 description 1
- 229960005183 paroxetine hydrochloride Drugs 0.000 description 1
- 150000003242 quaternary ammonium salts Chemical class 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000012421 spiking Methods 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000000825 ultraviolet detection Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/30—Control of physical parameters of the fluid carrier of temperature
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
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Abstract
A detection method for detecting the content of tetrabutylammonium bromide in levetiracetam is characterized by comprising the following steps: the method comprises the steps of adopting an ultra-high performance liquid chromatography to connect a triple quadrupole mass spectrometry (UPLC-MS/MS), taking an octadecylsilane chemically bonded silica filler column as a chromatographic column, taking acetonitrile as an organic phase, taking a volatile salt reagent solution as a water phase, mixing the acetonitrile and the volatile salt reagent solution as a mobile phase, eluting, and detecting by a mass spectrometry detector. The invention aims to provide a detection method for detecting the content of tetrabutylammonium bromide in levetiracetam, which has the advantages of good specificity, high sensitivity, simple sample preparation, rapidness and accuracy in detecting the content of tetrabutylammonium bromide in an organic medicine, thereby ensuring the quality control of the medicine and further ensuring the effectiveness and safety of the medicine.
Description
Technical Field
The invention relates to an analysis method for the content of tetrabutylammonium bromide in levetiracetam, in particular to a detection method for detecting the content of tetrabutylammonium bromide in levetiracetam.
Background
Controlling the level of impurities is an important method for ensuring the safety of pharmaceuticals. In the analysis of drug impurities, the harm of genotoxic impurities to human bodies is extremely large. Genotoxic impurities can damage cellular DNA directly or indirectly, producing mutagenic and carcinogenic substances. Genotoxic impurities are mainly derived from starting materials, intermediates, reagents and reaction byproducts in the synthesis of medicines, and meanwhile, in the preparation and storage links of medicines, the medicines are also likely to be degraded to generate the genotoxic impurities. In 2006, the european medicines agency issued a "genotoxic impurity limit guideline," and subsequently ICH and the U.S. food and drug administration also issued a control law for genotoxic impurities, which strengthened the regulation of genotoxic impurities.
Tetrabutylammonium bromide, also known as TBAB, is a quaternary ammonium salt and is widely applied to the synthesis process of various medicines, such as anti-infective medicines of ampicillin and sultamicin, antidiabetic medicines of dapagliflozin and the like. However, it has been found that tetrabutylammonium bromide has genotoxicity, and in paroxetine hydrochloride, the tetrabutylammonium bromide limit (25 ppm) needs to be strictly controlled to ensure the safety of the medicine in clinical application. The content limit of the tetrabutylammonium bromide is extremely low, and the need of an analysis method with high sensitivity and good specificity is very urgent.
Because tetrabutylammonium bromide has no ultraviolet absorption and is not easy to volatilize, conventional liquid chromatography ultraviolet detection and gas chromatography detection cannot be used, and currently, the detection methods mainly comprise an ion chromatography method and a bromide ion selective electrode method.
Bromide ion selective electrode method 1) poor sensitivity: very low levels of tetrabutylammonium bromide can be out of the linear range of the instrument; 2) the specificity is poor: is easily interfered by other bromide ion impurities in the medicine; 3) the variable factors are many: the temperature of the liquid to be detected, the stirring speed and the electrode variable factors in the detection process have great influence on the accuracy of the tetrabutylammonium bromide content.
Ion chromatography: 1) the sample treatment is complicated: ion chromatography detection requires that a sample is easily soluble in water, tetrabutylammonium bromide is easily soluble in water, but the solubility of an organic drug in water is poor, and very complex sample treatment is required to ensure analysis accuracy; 2) the ion chromatography is also used for determining the content of tetrabutylammonium bromide by detecting bromide ions in a solution and is influenced by other bromide ion impurities in the medicine.
Disclosure of Invention
The invention aims to provide a detection method for detecting the content of tetrabutylammonium bromide in levetiracetam, which has the advantages of good specificity, high sensitivity, simple sample preparation, rapidness and accuracy in detecting the content of tetrabutylammonium bromide in an organic medicine, thereby ensuring the quality control of the medicine and further ensuring the effectiveness and safety of the medicine.
The technical scheme of the invention is as follows:
a detection method for detecting the content of tetrabutylammonium bromide in levetiracetam is characterized by comprising the following steps: the method comprises the steps of adopting an ultra-high performance liquid chromatography to connect a triple quadrupole mass spectrometry (UPLC-MS/MS), taking an octadecylsilane chemically bonded silica filler column as a chromatographic column, taking acetonitrile as an organic phase, taking a volatile salt reagent solution as a water phase, mixing the acetonitrile and the volatile salt reagent solution as a mobile phase, eluting, and detecting by a mass spectrometry detector.
The concentration of the volatile salt reagent solution is 5-30 mmol/L; the volatile salt reagent is ammonium formate, a pH value regulator formic acid, and the content of the pH value regulator is 0.01-0.5%.
The elution volume ratio of the water phase is 18-22%.
The flow rate of the mobile phase is 0.27 ml/min-0.33 ml/min.
The temperature of the chromatographic column is 37-43 ℃.
The organic medicine is a western medicine, and the western medicine is an antiepileptic medicine.
The method of the present invention further comprises the steps of:
(1) taking a proper amount of organic medicine, adding acetonitrile, ultrasonically dissolving and diluting to prepare a solution containing 0.1-2 mg of the organic medicine per 1ml, preferably 0.2mg, passing through an organic filter membrane of 0.22 mu m, taking a subsequent filtrate, precisely transferring the subsequent filtrate into a measuring flask of 0.1 ml-10 ml, adding acetonitrile to dilute to a scale, and shaking up to be used as a sample solution. Taking a proper amount of tetrabutylammonium bromide reference substance, adding acetonitrile to dissolve and dilute the tetrabutylammonium bromide reference substance to prepare a solution containing 0.5-10 ng of the tetrabutylammonium bromide reference substance in every 1ml, and preferably 1ng of the tetrabutylammonium bromide reference substance as a reference substance solution;
(2) taking 1-10 mul of each of the test solution and the reference solution, preferably 5 mul, and injecting into a serial four-stage rod liquid chromatograph;
(3) the needle washing solution is acetonitrile, 50% acetonitrile solution, 50% methanol, methanol: water: formic acid (500: 500:1), preferably methanol: water: formic acid (500: 500: 1).
(4) The mass spectrum condition is that the Ion Source is ESI (+), the detection mode is a multi-reaction monitoring (MRM) mode, the spray voltage is 5500V, the Ion Source temperature is 550 ℃, Curtain Gas25 psi, Ion Source Gas 150 psi, Ion Source Gas 260 psi, and the residence time is 100 msec; other MRM parameters are shown in table 1.
TABLE 1 MRM parameter table for detecting tetrabutylammonium bromide content
(5) And calculating the content of tetrabutylammonium bromide in the organic medicine by using a peak area through an external standard method, and quantifying ions by using secondary fragments with the largest abundance.
The method has the following beneficial effects or advantages:
the method has strong specificity, can provide a quantitative result of the tetrabutylammonium bromide, and simultaneously has no interference on the determination by the solvent, the organic medicine and the impurities of the medicine;
the sensitivity is high, the minimum detection concentration of tetrabutylammonium bromide in the invention is 0.05ng/ml, and the detection of trace impurities can be ensured;
the durability is good, parameters such as column temperature, flow velocity and the like in the fine adjustment chromatographic condition fluctuate within the range of the invention, and the detection result is not obviously influenced;
the method has strong practicability, the tetrabutylammonium bromide can be determined within 5min, and the detection process is simple and convenient.
Drawings
FIG. 1 is a mass spectrum of a blank solvent;
FIG. 2 is a mass spectrum of a control solution;
FIG. 3 is a mass spectrum of a test solution;
FIG. 4 is a mass spectrum of a sample spiking solution.
FIG. 5 is a standard curve of tetrabutylammonium bromide.
Detailed Description
The present invention will be described in further detail with reference to the following examples:
as shown in fig. 1-5.
Example 1
The experimental conditions are as follows:
1) liquid chromatography conditions: a chromatographic column: waters HILIC 2.1 × 100mm,1.7 um. Mobile phase: a is 10.0mmol/L ammonium formate (0.1% formic acid) in water, B is acetonitrile, isocratic elution: 0-5.0 min, 80% of mobile phase B, flow rate: 0.3 ml/min; the column temperature is 40 ℃; sample introduction volume: 5 μ l, needle wash solution: methanol: water: formic acid (500: 500: 1).
2) Mass spectrum conditions: the ion source is ESI (+), the detection mode is a multi-reaction monitoring (MRM) mode, and the mass spectrum condition is as follows: the spraying voltage is 5500V, the Ion Source temperature is 550 ℃, Curtain Gas25 psi, Ion Source Gas 150 psi, Ion Source Gas 260 psi and the residence time is 100 msec; other MRM parameters are shown in table 2.
TABLE 2 MRM parameter table for detecting tetrabutylammonium bromide content
The experimental steps are as follows:
stock solution 1: accurately weighing 25mg of tetrabutylammonium bromide into a 25ml measuring flask, adding acetonitrile to dissolve, diluting to a scale, and shaking up. As stock solution 1 (1 mg/ml).
Stock solution 2: precisely transferring 0.1ml of stock solution into a measuring flask of 1-50 ml, adding acetonitrile to dissolve, diluting to a scale, and shaking up. As stock solution 2 (2. mu.g/ml).
Stock solution 3: precisely transferring 0.5ml of stock solution into a 2-10 ml measuring flask, adding acetonitrile to dissolve, diluting to a scale, and shaking up. As stock solution 3 (0.1. mu.g/ml).
Control solution (system suitability solution): and adding acetonitrile into a 3-10 ml measuring flask of 100 mu L stock solution for dissolution, diluting to a scale, and shaking up. As a control solution. (1 ng/ml).
Test solution: about 5.0mg of sample is taken, precisely weighed, dissolved in a 25ml measuring flask with acetonitrile and ultrasonically treated for 3min, and diluted to the scale. And (3) filtering the solution by using a 0.22 mu m organic filter membrane, precisely transferring the subsequent filtrate into a 0.1-10 ml measuring flask, adding acetonitrile to dilute the solution to a scale, and shaking the solution uniformly to serve as a test solution.
Adding a standard solution to a sample: sampling about 5.0mg of a sample, precisely weighing, dissolving and ultrasonically treating for 3min by using acetonitrile in a 25ml measuring flask, diluting to a scale, filtering by using a 0.22 mu m organic filter membrane, precisely transferring a subsequent filtrate into a 0.1ml to 10ml measuring flask, adding 100 mu L stock solution 3, diluting to the scale by using the acetonitrile, shaking uniformly, and taking the sample as a sample labeling solution.
The results show that: the blank solvent, the reference solution, the sample solution and the sample labeling solution are all free of interference, and the specification is good.
Example 2
The experimental conditions are as follows: the same as in example 1.
The experimental steps are as follows:
linear solution-10%: and adding acetonitrile into a 3-10 ml measuring flask of the 10 mu L stock solution for dissolution, and diluting until scales are uniformly shaken.
Linear solution-50%: and adding acetonitrile into a 3-10 ml measuring flask of 50 mu L stock solution for dissolution, and diluting until scales are uniformly shaken.
Linear solution-80%: and adding acetonitrile into a 3-10 ml measuring flask of 80 mu L stock solution for dissolution, and diluting until scales are uniformly shaken.
Linear solution-100%: and adding acetonitrile into a 3-10 ml measuring flask of 100 mu L stock solution for dissolution, and diluting until scales are uniformly shaken.
Linear solution-150%: and adding acetonitrile into a 3-10 ml measuring flask of 150 mu L stock solution for dissolution, and diluting until scales are uniformly shaken.
Linear solution-300%: and adding acetonitrile into a 3-10 ml measuring flask of 300 mu L stock solution for dissolution, and diluting until scales are uniformly shaken.
The experimental results are as follows: the peak area A was subjected to linear regression with the concentration C, and the test results are shown in Table 3. The concentration of tetrabutylammonium bromide is in a concentration range of 0.00010-0.00297 mu g/ml, the concentration and the peak area are in a linear relation, and the linear regression equation is as follows: y = 1,208,476,505.0000x-56,902.5736 (r = 0.997, n = 6), response factor RSD of 13.3%, Y-axis intercept at 5.0% of 100% response value. Meets the requirements.
TABLE 3 tetrabutylammonium bromide Standard Curve
Example 3
The experimental method comprises the following steps: the same as in example 1.
The experimental steps are as follows:
quantitative limiting solution: precisely moving and taking 35 mu l to 10mL of stock solution in a volumetric flask, adding acetonitrile to a constant volume to a scale, and shaking up. And precisely measuring 5 mu l of the measured signal, injecting the measured signal into a series-connection four-level rod liquid mass spectrometer, and comparing the measured signal with baseline noise to be used as a quantitative limit when the detected signal-to-noise ratio S/N is larger than or equal to 10.
Detection limit solution: precisely transferring the stock solution 32.5 mul to 10mL volumetric flask, adding acetonitrile to a constant volume to a scale, and shaking up. And precisely measuring 5 mu l of the measured signal, injecting the measured signal into a series four-level rod liquid chromatograph, and comparing the measured signal with baseline noise to be used as a detection limit when the detection signal-to-noise ratio S/N is larger than or equal to 3.
The experimental results are as follows: when the S/N is more than or equal to 3, the tetrabutylammonium bromide is 0.000025 mu g/ml. When the S/N is more than or equal to 10, the concentration of tetrabutylammonium bromide is 0.00005 mu g/ml, the detection requirement is met, in the sample injection precision test under the quantitative limit concentration term, the RSD of the peak area of tetrabutylammonium bromide is not more than 20%, and the RSD of the retention time is not more than 2%, so that the requirement is met.
TABLE 4 detection limit results
TABLE 5 quantitative limit results
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, and the preferred embodiments of the present invention are described in the above embodiments and the description, and are not intended to limit the present invention. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (7)
1. A detection method for detecting the content of tetrabutylammonium bromide in levetiracetam is characterized by comprising the following steps:
the method comprises the steps of adopting an ultra-high performance liquid chromatography to connect a triple quadrupole mass spectrometry (UPLC-MS/MS), taking an octadecylsilane chemically bonded silica filler column as a chromatographic column, taking acetonitrile as an organic phase, taking a volatile salt reagent solution as a water phase, mixing the acetonitrile and the volatile salt reagent solution as a mobile phase, eluting, and detecting by a mass spectrometry detector.
2. The method for detecting the content of tetrabutylammonium bromide in levetiracetam according to claim 1, wherein the concentration of the volatile salt reagent solution is 5-30 mmol/L; the volatile salt reagent is ammonium formate, a pH value regulator formic acid, and the content of the pH value regulator is 0.01-0.5%.
3. The method for detecting the content of tetrabutylammonium bromide in levetiracetam according to claim 1, wherein the elution volume ratio of the aqueous phase is 18-22%.
4. The method for detecting the content of tetrabutylammonium bromide in levetiracetam according to claim 1, wherein the flow rate of the mobile phase is 0.27ml/min to 0.33 ml/min.
5. The method for detecting the content of tetrabutylammonium bromide in levetiracetam according to claim 1, wherein the column temperature of the chromatographic column is 37-43 ℃.
6. The method for detecting the content of tetrabutylammonium bromide in levetiracetam according to claim 1, wherein the organic drug is a western drug, and the western drug is an antiepileptic drug.
7. The method for detecting the content of tetrabutylammonium bromide in levetiracetam according to claim 1, which is characterized by further comprising the following steps:
(1) taking a proper amount of organic medicine, adding acetonitrile, ultrasonically dissolving and diluting to prepare a solution containing 0.1-2 mg of the organic medicine per 1ml, preferably 0.2mg of the organic medicine, filtering with an organic filter membrane of 0.22 mu m, taking a subsequent filtrate, precisely transferring the subsequent filtrate into a measuring flask of 0.1 ml-10 ml, adding acetonitrile to dilute the solution to a scale, shaking up the solution to serve as a test solution, taking a proper amount of tetrabutylammonium bromide as a reference substance, adding acetonitrile to dissolve and dilute the solution to prepare a solution containing 0.5-10 ng of the organic medicine per 1ml, preferably 1ng of the organic medicine as a reference substance solution;
(2) taking 1-10 mul of each of the test solution and the reference solution, preferably 5 mul, and injecting into a serial four-stage rod liquid chromatograph;
(3) the needle washing solution is acetonitrile, 50% acetonitrile solution, 50% methanol, methanol: water: formic acid (500: 500:1), preferably methanol: water: formic acid (500: 500: 1);
(4) the mass spectrum condition is that the Ion Source is ESI (+), the detection mode is a multi-reaction monitoring (MRM) mode, the spray voltage is 5500V, the Ion Source temperature is 550 ℃, Curtain Gas25 psi, Ion Source Gas 150 psi, Ion Source Gas 260 psi, and the residence time is 100 msec; other MRM parameters are shown in table 1:
TABLE 1 MRM parameter table for detecting tetrabutylammonium bromide content
(5) And calculating the content of tetrabutylammonium bromide in the organic medicine by using a peak area through an external standard method, and quantifying ions by using secondary fragments with the largest abundance.
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|---|---|---|---|---|
| CN113624872A (en) * | 2021-08-03 | 2021-11-09 | 杭州微源检测技术有限公司 | Method for detecting content of 2-bromobutyric acid in etiracetam |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104678026A (en) * | 2015-03-19 | 2015-06-03 | 重庆医药工业研究院有限责任公司 | Method for measuring content of tetrabutylammonium bromide in organic drug |
| WO2019211799A1 (en) * | 2018-05-03 | 2019-11-07 | Institute Of Organic Chemistry And Biochemistry Ascr, V.V.I. | 2'3'-cyclic dinucleotide analogue comprising a cyclopentanyl modified nucleotide |
-
2020
- 2020-08-03 CN CN202010766302.5A patent/CN112034056A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104678026A (en) * | 2015-03-19 | 2015-06-03 | 重庆医药工业研究院有限责任公司 | Method for measuring content of tetrabutylammonium bromide in organic drug |
| WO2019211799A1 (en) * | 2018-05-03 | 2019-11-07 | Institute Of Organic Chemistry And Biochemistry Ascr, V.V.I. | 2'3'-cyclic dinucleotide analogue comprising a cyclopentanyl modified nucleotide |
Non-Patent Citations (2)
| Title |
|---|
| 刘继华等: "UPLC-MS/MS法测定F351中催化剂四丁基溴化铵的残留量", 《药物分析杂志》, no. 03, 31 March 2008 (2008-03-31) * |
| 刘继华等: "UPLC-MS/MS法测定硫辛酸中催化剂四丁基溴化铵的残留量", 《药物分析杂志》, no. 07, 31 July 2011 (2011-07-31), pages 1832 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113624872A (en) * | 2021-08-03 | 2021-11-09 | 杭州微源检测技术有限公司 | Method for detecting content of 2-bromobutyric acid in etiracetam |
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