CN115372528A - Detection method for simultaneously determining multiple impurities in nitrofurantoin - Google Patents
Detection method for simultaneously determining multiple impurities in nitrofurantoin Download PDFInfo
- Publication number
- CN115372528A CN115372528A CN202211141983.1A CN202211141983A CN115372528A CN 115372528 A CN115372528 A CN 115372528A CN 202211141983 A CN202211141983 A CN 202211141983A CN 115372528 A CN115372528 A CN 115372528A
- Authority
- CN
- China
- Prior art keywords
- impurity
- solution
- mobile phase
- reference substance
- nitrofurantoin
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000012535 impurity Substances 0.000 title claims abstract description 117
- NXFQHRVNIOXGAQ-YCRREMRBSA-N nitrofurantoin Chemical compound O1C([N+](=O)[O-])=CC=C1\C=N\N1C(=O)NC(=O)C1 NXFQHRVNIOXGAQ-YCRREMRBSA-N 0.000 title claims abstract description 55
- 229960000564 nitrofurantoin Drugs 0.000 title claims abstract description 55
- 238000001514 detection method Methods 0.000 title claims abstract description 29
- HYBBIBNJHNGZAN-UHFFFAOYSA-N furfural Chemical compound O=CC1=CC=CO1 HYBBIBNJHNGZAN-UHFFFAOYSA-N 0.000 claims abstract description 72
- 229960001907 nitrofurazone Drugs 0.000 claims abstract description 36
- IAIWVQXQOWNYOU-FPYGCLRLSA-N nitrofural Chemical compound NC(=O)N\N=C\C1=CC=C([N+]([O-])=O)O1 IAIWVQXQOWNYOU-FPYGCLRLSA-N 0.000 claims abstract description 35
- SXINBFXPADXIEY-UHFFFAOYSA-N 5-Nitrofurfural Chemical compound [O-][N+](=O)C1=CC=C(C=O)O1 SXINBFXPADXIEY-UHFFFAOYSA-N 0.000 claims abstract description 29
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims abstract description 29
- AAEQXEDPVFIFDK-UHFFFAOYSA-N 3-(4-fluorobenzoyl)-2-(2-methylpropanoyl)-n,3-diphenyloxirane-2-carboxamide Chemical compound C=1C=CC=CC=1NC(=O)C1(C(=O)C(C)C)OC1(C=1C=CC=CC=1)C(=O)C1=CC=C(F)C=C1 AAEQXEDPVFIFDK-UHFFFAOYSA-N 0.000 claims abstract description 27
- 238000000034 method Methods 0.000 claims abstract description 24
- 238000010828 elution Methods 0.000 claims abstract description 12
- 239000007788 liquid Substances 0.000 claims abstract description 11
- 239000000243 solution Substances 0.000 claims description 98
- 239000013558 reference substance Substances 0.000 claims description 96
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 87
- 239000002904 solvent Substances 0.000 claims description 33
- 238000005303 weighing Methods 0.000 claims description 31
- 238000007865 diluting Methods 0.000 claims description 18
- 239000011259 mixed solution Substances 0.000 claims description 13
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 12
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 12
- 239000012085 test solution Substances 0.000 claims description 10
- 230000003247 decreasing effect Effects 0.000 claims description 9
- 239000000945 filler Substances 0.000 claims description 6
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 6
- 239000000377 silicon dioxide Substances 0.000 claims description 6
- 239000008055 phosphate buffer solution Substances 0.000 claims description 5
- 239000012488 sample solution Substances 0.000 claims description 5
- FZAZWVSJBVAWTG-UHFFFAOYSA-N 3,4-diethyl-5-nitrofuran-2-carbaldehyde Chemical compound C(C)C=1C(=C(C=O)OC=1[N+](=O)[O-])CC FZAZWVSJBVAWTG-UHFFFAOYSA-N 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 239000003814 drug Substances 0.000 abstract description 7
- 230000035945 sensitivity Effects 0.000 abstract description 5
- 229940079593 drug Drugs 0.000 abstract description 4
- 150000001875 compounds Chemical class 0.000 abstract 1
- 238000012360 testing method Methods 0.000 description 14
- 238000011084 recovery Methods 0.000 description 11
- 238000000926 separation method Methods 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 238000012417 linear regression Methods 0.000 description 7
- 230000004044 response Effects 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- 238000002360 preparation method Methods 0.000 description 6
- 238000012795 verification Methods 0.000 description 6
- 239000011550 stock solution Substances 0.000 description 5
- 230000009286 beneficial effect Effects 0.000 description 4
- 239000012467 final product Substances 0.000 description 4
- 239000008363 phosphate buffer Substances 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 239000000463 material Substances 0.000 description 3
- 208000019206 urinary tract infection Diseases 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 2
- 231100000357 carcinogen Toxicity 0.000 description 2
- 239000003183 carcinogenic agent Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000004809 thin layer chromatography Methods 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 208000032840 Catheter-Related Infections Diseases 0.000 description 1
- 206010064687 Device related infection Diseases 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 208000035415 Reinfection Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000006482 condensation reaction Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 238000001310 location test Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000011471 prostatectomy Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
Images
Classifications
-
- 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
-
- 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
- G01N30/74—Optical detectors
Landscapes
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention relates to the technical field of drug detection, in particular to a detection method for simultaneously detecting various impurities in nitrofurantoin. The impurities are furacilin, furfural, 5-nitrofurfural diethyl ester, impurity D, impurity J and impurity K; the detection method is carried out by using a high performance liquid chromatograph, and the mobile phase consists of a mobile phase A and a mobile phase B and adopts gradient elution. The invention provides a detection method for simultaneously determining various impurities in nitrofurantoin by adopting a high performance liquid chromatograph according to the structural characteristics and the compound properties of a product. The method adopts gradient elution chromatographic conditions, can further simultaneously determine and separate the 6 impurities, can effectively separate the impurities from the nitrofurantoin, further improves the sensitivity and the accuracy of detection of each component by optimizing the conditions, meets the limit requirements of each impurity, and greatly reduces the detection cost and the operation difficulty.
Description
Technical Field
The invention relates to the technical field of drug detection, in particular to a detection method for simultaneously detecting various impurities in nitrofurantoin.
Background
Nitrofurantoin, marketed in the united states in 1953, was widely used in the 70-80 th 20 th century, and is a classic drug for the treatment of urinary tract infections. The medicine has no drug resistance problem in the application history of more than 60 years, can effectively treat acute lower urinary tract infection and chronic urinary tract infection and prevent catheter-related infection, can be used as a basic medicine for preventing transurethral prostatectomy infection, and can prevent re-infection. Its action is mainly due to its high concentration in urine and low concentration in plasma, effective coverage of gram-positive and negative bacteria, and few resistant strains are produced, superior to other new drugs.
Furacilin is a by-product in nitrofurantoin synthesis, has high toxicity, and is controlled by an HPLC method in the 2020 edition of Chinese pharmacopoeia and the quality standard of the USP-NF 2021 nitrofurantoin of United states pharmacopoeia, with the limit of 0.01 percent.
5-nitrofurfural diethyl ester is a starting material for synthesizing nitrofurantoin, is brought in the last step of condensation reaction and should be controlled in the nitrofurantoin, currently, only United states pharmacopoeia USP-NF 2021 in pharmacopoeias of various countries adopts thin layer chromatography for control, but the method sensitivity is lower, and the limit is not more than 1.0%.
Furfural is used as a starting material (5-nitrofurfural diethyl ester) for synthesis. 2017, 10, 27, a list of carcinogens published by the world health organization international cancer research institute, and furfural is in the list of class 3 carcinogens. TD of Furfural based on CPDB database 50 683 mg/kg/day (rat) and 197 mg/kg/day (mouse), the lower mouse values were used to calculate the acceptable human intake of 197 μ g/day and the maximum daily dose of nitrofurantion of 400mg, which calculated the limit of control of this impurity in nitrofurantion to 0.05%, whereas furfural was not controlled in the pharmacopoeias of various countries.
Other related substances possibly exist in nitrofurantoin, the U.S. pharmacopoeia is not controlled, the Chinese pharmacopoeia, the European pharmacopoeia and the British pharmacopoeia are all controlled by only adopting a thin layer chromatography, the method has low sensitivity, and the control requirement of the ICHQ3A cannot be met. ICH Q3A requires that for raw material drugs with maximum daily dose of 2g or less, the identification limit of impurities is set to 0.10% or 1.0mg (lower limit) taken per day, the maximum daily dose of nitrofurantoin is less than 2g, and the unknown single impurity control limit is set to be below 0.10%.
In the prior art, only furacilin and furfural which are impurities in nitrofurantoin are detected, and the furacilin and the furfural need to be detected respectively under different chromatographic conditions, so that the problems of high detection cost, long time consumption and low sensitivity are caused to pharmaceutical enterprises, and the method is not beneficial to popularization and use.
Disclosure of Invention
The present invention is directed to solving at least one of the problems of the prior art, and therefore an aspect of the present invention is to provide a detection method for simultaneously detecting a plurality of impurities in nitrofurantoin, such as nitrofurazone, furfural, diethyl 5-nitrofuraldehyde, impurity D, impurity J, and impurity K; the detection method is carried out by using a high performance liquid chromatograph, and the mobile phase consists of a mobile phase A and a mobile phase B and adopts gradient elution. The chemical structural formula of the impurity is as follows:
the invention discovers that when the high performance liquid chromatography is adopted to detect nitrofurazone, 5-nitrofurfural diethyl ester, furfural and other related substances in nitrofurantoin, the selection of a mobile phase has great influence on the separation of impurities, and when the mobile phase A and the mobile phase B with fixed proportion are adopted to detect, the impurities are difficult to be well separated; through a great deal of attempts, the nitrofurantoin, nitrofurazone, 5-nitrofuraldehyde diethyl ester, furfural and other impurities can be effectively separated by adopting the elution mode, and the running time is shortened.
Preferably, the specific operation steps of the detection method are as follows:
s1, sample solution: taking nitrofurantoin, precisely weighing, adding a solvent for dissolving, and quantitatively diluting to prepare a test solution;
s2, comparison solution: precisely measuring a test solution, and quantitatively diluting with a solvent to obtain a nitrofurantoin control solution;
s3, mixing solution: dissolving nitrofurantoin reference substance in solvent, precisely adding nitrofurazone reference substance, furfural reference substance, 5-nitrofuraldehyde diethyl ester reference substance, impurity D reference substance, impurity J reference substance, and impurity K reference substance, and quantitatively diluting to obtain mixed solution;
s4, positioning solution of each impurity: taking a furacilin reference substance, precisely weighing, and dissolving with a solvent to prepare a furacilin positioning solution; taking a furfural reference substance, precisely weighing, and adding a solvent to dissolve to prepare a furfural positioning solution; taking a 5-nitrofurfural diethyl ester reference substance, precisely weighing, and adding a solvent to dissolve to prepare a 5-nitrofurfural diethyl ester positioning solution; precisely weighing an impurity D reference substance, and dissolving the impurity D reference substance by adding a solvent to prepare an impurity D positioning solution; precisely weighing impurity J reference substance, and dissolving with solvent to obtain impurity J positioning solution; precisely weighing an impurity K reference substance, and dissolving the impurity K reference substance in a solvent to prepare an impurity K positioning solution;
s5, adjusting an instrument: octadecylsilane chemically bonded silica is used as a filling agent for a chromatographic column, a mobile phase A and a mobile phase B are used for regulating the column temperature, the flow velocity of the mobile phase and the detection wavelength, and gradient elution is adopted;
s6, determination: and precisely measuring each solution prepared from S2 to S4, respectively injecting the solution into a liquid chromatograph, and recording a chromatogram.
Preferably, the concentration of the sample solution in S1 is 0.8-1.2 mg/ml.
Preferably, the concentration of the control solution in S2 is 0.08-0.12 μ g/ml.
Preferably, the concentration of the nitrofurantoin reference substance in the mixed solution in S3 is 0.8-1.2 mg/ml, the concentration of the nitrofurazon reference substance is 0.08-0.12 mu g/ml, the concentration of the furfural reference substance is 0.4-0.6 mu g/ml, and the concentrations of the 5-nitrofurfural diethyl ester reference substance, the impurity D reference substance, the impurity J reference substance and the impurity K reference substance are all 0.8-1.2 mu g/ml.
Preferably, the concentration of each impurity positioning solution nitrofurazone positioning solution in the S4 is 0.08-0.12 mu g/ml; the concentration of the furfural positioning solution is 0.4-0.6 mu g/ml; the concentration of the 5-nitrofurfural diethyl ester positioning solution is 0.8-1.2 mu g/ml; the concentration of the impurity D positioning solution is 0.8-1.2 mug/ml; the concentration of the impurity J positioning solution is 0.8-1.2 mug/ml; the concentration of the impurity K positioning solution is 0.8-1.2 mu g/ml.
Preferably, the solvent is 50% acetonitrile.
Preferably, the chromatographic column in S5 uses octadecylsilane chemically bonded silica as a filler, and the specification is Inertsil ODS-3, 250mm multiplied by 4.6mm,5 μm.
Preferably, the mobile phase A is phosphate buffer solution, the concentration of the phosphate buffer solution is 0.05mol/L, the pH value is adjusted to 7.1 +/-0.1 by using sodium hydroxide solution, and the mobile phase B is 100% acetonitrile; adjusting the column temperature to 25-35 ℃, adjusting the flow rate of the mobile phase to 0.9-1.1 ml/min, adjusting the detection wavelength to 375nm to detect nitrofurantoin, nitrofurazone, D and K, and 304nm to detect diethyl 5-nitrofuraldehyde, furfural and J. The pH value of the phosphate buffer has great influence on the separation degree of the nitrofurantoin and the nitrofurazone, and when the pH value of the phosphate buffer is 7.1 +/-0.1, the separation effect is better; not only can ensure that the separation degree of the nitrofurantoin and the nitrofurazone reaches more than 4.0, but also can ensure that other impurities are effectively separated. Because the difference of ultraviolet absorption wavelengths of all impurities in the nitrofurantoin is large, all the impurities are difficult to be detected at the same wavelength, and the invention can detect 6 impurities by using the wavelengths of 375nm and 304 nm.
Preferably, the gradient elution is adopted, the total volume of the mobile phase is 100%,
in 0-4 min, the volume of the mobile phase A is 98 percent, and the volume of the mobile phase B is 2 percent;
in the 4 th to 30 th min, the volume of the mobile phase A is decreased from 100% to 80%, and the volume of the mobile phase B is increased from 2% to 20%;
at 30-60 min, the volume of the mobile phase A is decreased from 80% to 45%, and the volume of the mobile phase B is increased from 20% to 55%;
at the time of 60-65 min, the volume of the mobile phase A is 45%, and the volume of the mobile phase B is 55%;
at 65-66 min, the volume of the mobile phase A is increased from 45% to 98%, and the volume of the mobile phase B is decreased from 55% to 2%;
at 66-75 min, the volume of the mobile phase A is 98%, and the volume of the mobile phase B is 2%.
The invention has the following beneficial effects:
the invention provides a detection method for simultaneously determining various impurities in nitrofurantoin by using a high performance liquid chromatograph according to the structural characteristics and the properties of a product. The method adopts gradient elution chromatographic conditions, can further simultaneously determine and separate the 6 impurities, can effectively separate the impurities from the nitrofurantoin, further improves the sensitivity and the accuracy of detection of each component by optimizing the conditions, meets the limit requirements of each impurity, and greatly reduces the detection cost and the operation difficulty.
The method has the advantages of simple equipment, high analysis speed, good specificity, low detection cost, high accuracy, and capability of quickly, simply and accurately controlling the amount of impurities and reducing the safety risk of the medicine. The method has strong specificity and high repeatability, is convenient for quality detection and impurity monitoring of the nitrofurantoin, is beneficial to popularization of the nitrofurantoin impurity detection method, and is beneficial to safe popularization and application of the nitrofurantoin.
Additional aspects and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention.
Drawings
The above and/or additional aspects and advantages of the present invention will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:
FIG. 1 is a chromatogram of a mixed solution at a wavelength of 375nm in a first embodiment of the present invention;
FIG. 2 is a chromatogram of a mixed solution at a wavelength of 304nm in the first embodiment of the present invention.
Detailed Description
In order that the above objects, features and advantages of the present invention can be more clearly understood, the present invention will be described in further detail with reference to the accompanying drawings and specific embodiments. It should be noted that the embodiments and features of the embodiments of the present application may be combined with each other without conflict.
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, however, the present invention may be practiced otherwise than as specifically described and thus the scope of the present invention is not limited by the specific embodiments disclosed below.
The present example provides a method for simultaneously measuring a plurality of impurities in nitrofurantoin by using a high performance liquid chromatograph, which comprises the following steps:
a chromatographic column: octadecylsilane chemically bonded silica is used as a filler, and the specification of a chromatographic column is Inertsil ODS-3, 250mm multiplied by 4.6mm,5 mu m; mobile phase A: phosphate buffer (the concentration of the phosphate buffer is 0.05mol/L, and the pH value is adjusted to 7.1 +/-0.1 by using sodium hydroxide solution); mobile phase B:100% acetonitrile; flow rate: 0.9-1.1 ml/min; column temperature: 25 to 35 ℃; detection wavelength: 375nm,304nm; gradient elution is used, and the total volume of the mobile phase is 100%.
In 0-4 min, the volume of the mobile phase A is 98 percent, and the volume of the mobile phase B is 2 percent;
in 4-30 min, the volume of the mobile phase A is decreased from 100% to 80%, and the volume of the mobile phase B is increased from 2% to 20%;
at 30-60 min, the volume of the mobile phase A is decreased from 80% to 45%, and the volume of the mobile phase B is increased from 20% to 55%;
at 60-65 min, the volume of the mobile phase A is 45%, and the volume of the mobile phase B is 55%;
at 65-66 min, the volume of the mobile phase A is increased from 45% to 98%, and the volume of the mobile phase B is decreased from 55% to 2%;
at 66-75 min, the volume of the mobile phase A is 98%, and the volume of the mobile phase B is 2%.
Test solution: the nitrofurantoin is precisely weighed, dissolved by a solvent (50% acetonitrile solution) and quantitatively diluted by water to prepare a test sample solution of 1 mg/ml.
Control solution: the sample solution was precisely measured and quantitatively diluted with a solvent (50% acetonitrile solution) to prepare a 1. Mu.g/ml nitrofurantoin control solution.
Mixing the solution: dissolving nitrofurantoin reference substance in 50% acetonitrile solution, precisely adding nitrofurazone reference substance, furfural reference substance, 5-nitrofurfural diethyl ester reference substance, impurity D reference substance, impurity J reference substance, and impurity K reference substance, and quantitatively diluting to obtain mixed solution; wherein the concentration of the nitrofurantoin reference substance is 1mg/ml, the concentration of the nitrofurazone reference substance is 0.1 mu g/ml, the concentration of the furfural reference substance is 0.5 mu g/ml, and the concentrations of the 5-nitrofurfural diethyl ester reference substance, the impurity D reference substance, the impurity J reference substance and the impurity K reference substance are respectively 1 mu g/ml.
Each impurity localization solution: taking a furacilin reference substance, precisely weighing, and dissolving with a solvent (50% acetonitrile solution) to obtain a furacilin positioning solution with the concentration of 0.1 mu g/ml;
taking a furfural reference substance, precisely weighing, adding a solvent (50% acetonitrile solution) to dissolve to prepare a furfural positioning solution with the concentration of 0.5 mu g/ml;
taking a 5-nitrofurfural diethyl ester reference substance, precisely weighing, adding a solvent (50% acetonitrile solution) to dissolve to prepare a 5-nitrofurfural diethyl ester positioning solution with the concentration of 1 mu g/ml;
precisely weighing an impurity D reference substance, and dissolving with a solvent (50% acetonitrile solution) to obtain an impurity D positioning solution with the concentration of 1 microgram/ml;
precisely weighing impurity J reference substance, and dissolving with solvent (50% acetonitrile solution) to obtain impurity J positioning solution with concentration of 1 μ g/ml;
taking an impurity K reference substance, precisely weighing, and adding a solvent (50% acetonitrile solution) to dissolve to prepare an impurity K positioning solution with the concentration of 1 mu g/ml.
The specific preparation process of each solution and the experimental verification process and results of the method of the invention are shown in experimental examples one to four.
Example one
Specificity test
Preparation of a test solution: the nitrofurantoin is precisely weighed to be about 20mg, placed in a 20ml measuring flask, dissolved by adding 10ml of 50 percent acetonitrile, diluted to the scale with water and shaken up to be used as a test solution.
Preparation of control solution: a suitable amount of the test solution was measured precisely, and diluted with a solvent to give a solution containing 1. Mu.g of nitrofurantoin per 1ml, which was used as a control solution.
Preparation of mixed solution: taking about 20mg of nitrofurantoin reference substance, adding 10ml of acetonitrile for dissolving, precisely adding a proper amount of nitrofurazone reference substance, furfural reference substance, 5-nitrofuraldehyde diethyl ester reference substance, impurity D reference substance, impurity J reference substance and impurity K reference substance, adding water for diluting to prepare a solution containing 1mg of nitrofurantoin reference substance, 0.1 mu g of nitrofurazone reference substance, 0.5 mu g of furfural reference substance, and 1 mu g of 5-nitrofuraldehyde diethyl ester reference substance, impurity D reference substance, impurity J reference substance and impurity K reference substance in each 1ml, and taking the solution as a mixed solution.
Preparing an impurity positioning solution: taking a proper amount of furacilin reference substances, precisely weighing, adding 50% acetonitrile for dissolving and diluting to obtain a solution of 0.1 mu g/ml, and taking the solution as a furacilin positioning solution; taking a proper amount of furfural reference substance, precisely weighing, adding 50% acetonitrile for dissolving and diluting to obtain a solution of 0.5 mu g/ml, and taking the solution as a furfural positioning solution; taking a proper amount of a 5-nitrofurfural diethyl ester reference substance, precisely weighing, adding 50% acetonitrile to dissolve and dilute to obtain a solution with the concentration of 1 mu g/ml, and taking the solution as a 5-nitrofurfural diethyl ester positioning solution; taking appropriate amount of impurity D reference substance, impurity J reference substance and impurity K reference substance, precisely weighing, dissolving with 50% acetonitrile, and diluting to obtain 1 μ g/ml solution as impurity D, impurity J and impurity K positioning solution.
And (3) determination: octadecylsilane chemically bonded silica is used as a filler for a chromatographic column, a mobile phase A is phosphate buffer solution (6.8 g of monopotassium phosphate is dissolved by adding 500ml of water, the pH value is adjusted to 7.1 +/-0.1 by using 1mol/L sodium hydroxide solution), a mobile phase B is 50% acetonitrile, the column temperature is 30 ℃, the flow rate is 1ml per minute, the detection wavelength is 375nm and 304nm, and gradient elution is adopted.
Precisely measuring the prepared control solution, the mixed solution and each impurity positioning solution by 10 μ l, injecting into a high performance liquid chromatograph, and recording chromatogram. The results are shown in Table 1, and the chromatograms of the mixed solutions are shown in FIGS. 1 and 2.
TABLE 1 results of specificity-location test
And (4) conclusion: the solvent does not interfere the determination of known impurities in the test solution, the separation among the impurities, the impurities and the main peak is good, and the number of theoretical plates meets the requirements of the determination of related substances.
Example two
Linear and range test
Preparation of linear solution: taking appropriate amounts of a nitrofurantoin reference substance, a nitrofurazone reference substance, a 5-nitrofuraldehyde diethyl ester reference substance, an impurity D reference substance, a furfural reference substance, an impurity J reference substance and an impurity K reference substance, precisely weighing appropriate amounts, dissolving with acetonitrile, quantitatively diluting to obtain solutions each containing 200 mu g of impurities per 1ml, taking the solutions as storage solutions, precisely weighing appropriate amounts, and diluting with a solvent (50% acetonitrile solution) to obtain a series of linear solutions.
Precisely measuring 10 μ l of each solution, injecting into a liquid chromatograph, and recording chromatogram; the results are shown in Table 2.
TABLE 2 results of the Linear and Range tests
And (4) conclusion: (1) The linear relation of nitrofurantoin in the concentration range of 0.032-2.107 mug/ml (corresponding to 0.003% -0.20% of the detected concentration) is good, the linear regression equation is y =46625.0590x +366.5408, r =0.9998, the percentage of the Y-axis intercept corresponding to 100% concentration response value is 0.7%.
(2) Furacilin: the linear relation is good in the concentration range of 0.020-0.196 mug/ml (corresponding to 0.002% -0.020% of the detected concentration), the linear regression equation is y =51453.0467x +16.0732, r =0.9997, the percentage of the Y-axis intercept corresponding to 100% of the concentration response value is 0.3%.
(3) Impurity D: the linear relation is good in the concentration range of 0.029-1.960 mu g/ml (corresponding to 0.003% -0.20% of the detected concentration), the linear regression equation is y =38339.0547x +345.4003, r =0.9998, and the percentage of the Y-axis intercept corresponding to 100% of the concentration response value is 0.9%.
(4) Impurity K: the linear relation is good in the concentration range of 0.030-2.025 mu g/ml (corresponding to 0.003% -0.2% of the detected concentration), the linear regression equation is y =38034.2562x +163.6797, r =0.9998, and the percentage of the Y-axis intercept corresponding to 100% of the concentration response value is 0.4%.
(5) 5-nitrofurfural diethyl ester: the linear relation is good in the concentration range of 0.050-1.997 mug/ml (corresponding to 0.005% -0.20% of the detected concentration), the linear regression equation is y =29072.9433x +94.8157, r =1.0000, and the percentage of the Y-axis intercept corresponding to 100% concentration response value is 0.3%.
(5) And (3) furfural: the linear relation is good in the concentration range of 0.099-0.990 mug/ml (corresponding to 0.01% -0.1% of the detected concentration), the linear regression equation is y =10382.3386x +48.6341, r =0.9997, the percentage of the Y-axis intercept corresponding to 100% concentration response value is 0.9%.
(7) Impurity J: the linear relation is good in the concentration range of 0.021-2.061 mug/ml (corresponding to 0.002% -0.2% of the detected concentration), the linear regression equation is y =74858.6593x +508.6866, r =0.9998, and the percentage of the Y-axis intercept corresponding to 100% of the concentration response value is 0.6%.
EXAMPLE III
Recovery test
Impurity reference substance: taking a proper amount of furacilin, 5-nitrofurfural diethyl ester, an impurity D, furfural, an impurity J and an impurity K as reference substances, precisely weighing, dissolving with acetonitrile, and quantitatively diluting with a solvent (50% acetonitrile solution) to prepare a solution containing 1 mu g/ml of furacilin, 5 mu g/ml of furfural and 10 mu g/ml of other impurities in each 1ml of solution as an impurity reference substance stock solution. Precisely measuring 2ml of the reference stock solution, placing in a 20ml measuring flask, diluting with solvent to scale, and shaking; 2 parts are prepared in parallel.
Preparation of an accuracy solution:
50% accuracy solution: precisely weighing about 20mg of a test sample, placing in a 20ml measuring flask, adding about 10ml of acetonitrile for dissolving, adding 1ml of impurity reference substance stock solution, adding water for diluting to scale, and shaking up to obtain the final product; 3 parts are prepared in parallel.
100% accuracy solution: precisely weighing about 20mg of a test sample, placing in a 20ml measuring flask, adding about 10ml of acetonitrile for dissolving, adding 2ml of impurity reference substance stock solution, adding water for diluting to scale, and shaking up to obtain the final product; 6 parts are prepared in parallel.
150% accuracy solution: precisely weighing about 20mg of a test sample, placing in a 20ml measuring flask, adding about 10ml of acetonitrile for dissolving, adding 3ml of impurity reference substance stock solution, adding water for diluting to scale, and shaking up to obtain the final product; 3 parts are prepared in parallel.
Preparing a background solution:
taking about 20mg of a test sample, precisely weighing, placing in a 20ml measuring flask, adding about 10ml of acetonitrile to dissolve, adding water to dilute to a scale, and shaking up to obtain the final product.
The solutions were measured precisely at 10. Mu.l each and injected into a liquid chromatograph, and the results are shown in tables 3 to 8.
TABLE 3 verification of related substance methods-Furacilin recovery results
TABLE 4 relevant materials method validation-results on recovery of impurity D
TABLE 5 verification of related materials methods-results on recovery of impurity K
TABLE 6 verification of 5-Nitro-furfural diethyl ester recovery results by related substance methods
TABLE 7 verification of related materials methods-Furfural recovery results
TABLE 8 verification of related substances methods-results on recovery of impurity J
And (4) conclusion: the result of the impurity recovery rate test shows that the recovery rates of 12 accuracy samples of nitrofurazone, an impurity D, an impurity K, 5-nitrofurfural diethyl ester, furfural and an impurity J are all 80-120%, the average recovery rates are 104.54%, 104.14%, 101.36%, 100.14%, 105.32% and 101.51%, and the table 3-9 shows that the recovery rates of the impurities meet the measurement requirements of the product on the known impurities, and the method is good in accuracy.
Example four
Durability test
Taking the mixed solution in the first experimental example, and respectively carrying out column temperature of 25 ℃ and 35 ℃; the flow rate is 0.9ml/min and 1.1ml/min; the results of the tests carried out at pH7.0 and pH7.2 for the mobile phase are shown in tables 9 to 10.
TABLE 9 durability test results (375 nm mixed solution-degree of separation)
TABLE 10 durability test results (304 nm mixed solution-degree of separation)
And (4) conclusion: under the condition of fine-tuning chromatography, the separation degrees among impurities, impurities and main peaks in the mixed solution meet the requirements. Indicating that the method is of good durability.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and it is obvious to those skilled in the art that various modifications and variations can be made in the present invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A detection method for simultaneously determining multiple impurities in nitrofurantoin is characterized by comprising the following steps: the impurities are furacilin, furfural, 5-nitrofurfural diethyl ester, impurity D, impurity J and impurity K; the detection method is carried out by using a high performance liquid chromatograph, and the mobile phase consists of a mobile phase A and a mobile phase B and adopts gradient elution.
2. The method for simultaneously detecting a plurality of impurities in nitrofurantoin according to claim 1, wherein: the detection method comprises the following specific operation steps:
s1, sample solution: taking nitrofurantoin, precisely weighing, adding a solvent for dissolving, and quantitatively diluting to prepare a test solution;
s2, comparison solution: precisely measuring a test solution, and quantitatively diluting with a solvent to obtain a nitrofurantoin control solution;
s3, mixing solution: dissolving nitrofurantoin reference substance in solvent, precisely adding nitrofurazone reference substance, furfural reference substance, 5-nitrofuraldehyde diethyl ester reference substance, impurity D reference substance, impurity J reference substance, and impurity K reference substance, and quantitatively diluting to obtain mixed solution;
s4, positioning each impurity: taking a furacilin reference substance, precisely weighing, and dissolving with a solvent to prepare a furacilin positioning solution; taking a furfural reference substance, precisely weighing, and adding a solvent to dissolve to prepare a furfural positioning solution; taking a 5-nitrofurfural diethyl ester reference substance, precisely weighing, and dissolving with a solvent to prepare a 5-nitrofurfural diethyl ester positioning solution; precisely weighing the impurity D reference substance, and dissolving with a solvent to obtain an impurity D positioning solution; precisely weighing impurity J reference substance, and dissolving with solvent to obtain impurity J positioning solution; precisely weighing an impurity K reference substance, and dissolving the impurity K reference substance in a solvent to prepare an impurity K positioning solution;
s5, adjusting an instrument: octadecylsilane chemically bonded silica is used as a filling agent for a chromatographic column, a mobile phase A and a mobile phase B are used for adjusting the column temperature, the flow velocity and the detection wavelength of the mobile phase, and gradient elution is adopted;
s6, determination: and precisely measuring each solution prepared from S2 to S4, respectively injecting the solution into a liquid chromatograph, and recording a chromatogram.
3. The method for simultaneously detecting a plurality of impurities in nitrofurantoin according to claim 2, wherein: the concentration of the test solution in the S1 is 0.8-1.2 mg/ml.
4. The method for simultaneously detecting a plurality of impurities in nitrofurantoin according to claim 2, wherein: the concentration of the control solution in the S2 is 0.08-0.12 mu g/ml.
5. The method for simultaneously detecting a plurality of impurities in nitrofurantoin according to claim 2, wherein: the concentration of the nitrofurazone reference substance in the mixed solution in the S3 is 0.8-1.2 mg/ml, the concentration of the nitrofurazone reference substance is 0.08-0.12 mu g/ml, the concentration of the furfural reference substance is 0.4-0.6 mu g/ml, and the concentrations of the 5-nitrofurfural diethyl ester reference substance, the impurity D reference substance, the impurity J reference substance and the impurity K reference substance are all 0.8-1.2 mu g/ml.
6. The method for simultaneously detecting a plurality of impurities in nitrofurantoin according to claim 2, wherein: the concentration of each impurity positioning solution furacilin positioning solution in the S4 is 0.08-0.12 mu g/ml; the concentration of the furfural positioning solution is 0.4-0.6 mu g/ml; the concentration of the 5-nitrofurfural diethyl ester positioning solution is 0.8-1.2 mu g/ml; the concentration of the impurity D positioning solution is 0.8-1.2 mug/ml; the concentration of the impurity J positioning solution is 0.8-1.2 mu g/ml; the concentration of the impurity K positioning solution is 0.8-1.2 mu g/ml.
7. The method for simultaneously detecting a plurality of impurities in nitrofurantoin according to any of claims 2 to 6, wherein: the solvent is 50% acetonitrile.
8. The method for simultaneously detecting a plurality of impurities in nitrofurantoin according to claim 2, wherein: the chromatographic column in the S5 uses octadecylsilane chemically bonded silica as a filler, and the specification is Inertsil ODS-3, 250mm multiplied by 4.6mm,5 mu m.
9. The method for simultaneously detecting a plurality of impurities in nitrofurantoin according to claim 2 or 8, wherein: the mobile phase A is phosphate buffer solution, the concentration of the phosphate buffer solution is 0.05mol/L, the pH value is adjusted to 7.1 +/-0.1 by using sodium hydroxide solution, and the mobile phase B is 100% acetonitrile; adjusting the column temperature to 25-35 ℃, adjusting the flow rate of the mobile phase to 0.9-1.1 ml/min, adjusting the detection wavelength to 375nm to detect nitrofurantoin, nitrofurazone, D and K, and 304nm to detect diethyl 5-nitrofuraldehyde, furfural and J.
10. The method for simultaneously detecting a plurality of impurities in nitrofurantoin according to claim 2, wherein: the gradient elution is adopted, the total volume of the mobile phase is 100 percent,
in 0-4 min, the volume of the mobile phase A is 98 percent, and the volume of the mobile phase B is 2 percent;
in the 4 th to 30 th min, the volume of the mobile phase A is decreased from 100% to 80%, and the volume of the mobile phase B is increased from 2% to 20%;
at 30-60 min, the volume of the mobile phase A is decreased from 80% to 45%, and the volume of the mobile phase B is increased from 20% to 55%;
at 60-65 min, the volume of the mobile phase A is 45%, and the volume of the mobile phase B is 55%;
at 65-66 min, the volume of the mobile phase A is increased from 45% to 98%, and the volume of the mobile phase B is decreased from 55% to 2%;
at 66-75 min, the volume of the mobile phase A is 98%, and the volume of the mobile phase B is 2%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211141983.1A CN115372528B (en) | 2022-09-20 | 2022-09-20 | Detection method for simultaneously measuring various impurities in nitrofurantoin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211141983.1A CN115372528B (en) | 2022-09-20 | 2022-09-20 | Detection method for simultaneously measuring various impurities in nitrofurantoin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115372528A true CN115372528A (en) | 2022-11-22 |
| CN115372528B CN115372528B (en) | 2023-06-23 |
Family
ID=84070771
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211141983.1A Active CN115372528B (en) | 2022-09-20 | 2022-09-20 | Detection method for simultaneously measuring various impurities in nitrofurantoin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115372528B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102830192A (en) * | 2012-08-06 | 2012-12-19 | 中国水产科学研究院东海水产研究所 | Method for simultaneously detecting nitrofurans raw drug residue in aquatic product |
| CN103926340A (en) * | 2014-04-04 | 2014-07-16 | 中国检验检疫科学研究院 | Method for measuring nitrofuran antibiotics in cosmetics |
| CN107655984A (en) * | 2017-08-03 | 2018-02-02 | 昌邑市检验检测中心 | Nitrofuran metabolites method for detecting residue in a kind of poultry |
| CN108008038A (en) * | 2017-11-26 | 2018-05-08 | 杭州市食品药品检验研究院 | Measure four kinds of remaining methods of Nitrofuran metabolites metabolite in gelatin type traditional Chinese medicine |
-
2022
- 2022-09-20 CN CN202211141983.1A patent/CN115372528B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102830192A (en) * | 2012-08-06 | 2012-12-19 | 中国水产科学研究院东海水产研究所 | Method for simultaneously detecting nitrofurans raw drug residue in aquatic product |
| CN103926340A (en) * | 2014-04-04 | 2014-07-16 | 中国检验检疫科学研究院 | Method for measuring nitrofuran antibiotics in cosmetics |
| CN107655984A (en) * | 2017-08-03 | 2018-02-02 | 昌邑市检验检测中心 | Nitrofuran metabolites method for detecting residue in a kind of poultry |
| CN108008038A (en) * | 2017-11-26 | 2018-05-08 | 杭州市食品药品检验研究院 | Measure four kinds of remaining methods of Nitrofuran metabolites metabolite in gelatin type traditional Chinese medicine |
Non-Patent Citations (2)
| Title |
|---|
| P. VIN˜AS ET AL.: "Analysis of Nitrofuran Residues in Animal Feed Using Liquid Chromatography and Photodiode-Array Detection", 《CHROMATOGRAPHIA》, vol. 65 * |
| 徐长根 等: "呋喃妥因肠溶片中有关物质检测及结构分析", 《药物分析杂志》, vol. 32, no. 12 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115372528B (en) | 2023-06-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107315059B (en) | The content assaying method of rifampin and its impurity in a kind of rifampicin capsules | |
| CN107064350B (en) | Method for detecting suspected genotoxic impurity of tofacitinib citrate | |
| CN105301126B (en) | Method for analyzing topiroxostat-related substances | |
| CN108061767A (en) | The method of HPLC method separation determination Rivaroxaban intermediates and its related impurities | |
| CN111024831B (en) | Method for separating moxifloxacin hydrochloride and impurities thereof by high performance liquid chromatography | |
| CN109580825B (en) | Method for detecting p-toluenesulfonate substances in racecadotril | |
| CN113049687A (en) | Method for detecting related substances of ambroxol hydrochloride raw material and injection | |
| CN113933399B (en) | Method for separating and detecting rifampicin and related impurities in rifampicin for injection | |
| CN108414636A (en) | A kind of detection method of the cinnarizine in relation to substance | |
| CN106706769B (en) | Separation and determination method of empagliflozin and optical isomer thereof | |
| CN115372528B (en) | Detection method for simultaneously measuring various impurities in nitrofurantoin | |
| CN113820409B (en) | Method for detecting related substances in mother nucleus of moxifloxacin | |
| CN107014943B (en) | A kind of detection method of ibrutinib enantiomer | |
| CN111380991B (en) | Method for detecting content of degradation impurities in vitamin C medicament | |
| CN111521693B (en) | Method for detecting isosorbide mononitrate | |
| CN115791989A (en) | Separation and detection method for impurities in cefoperazone sodium or preparation thereof | |
| CN107870210A (en) | The method for determining Li Gelieting contents in Li Gelieting bulk drugs | |
| CN112557558B (en) | Method for detecting SCH59566 impurity content in ezetimibe simvastatin tablets | |
| CN106442793B (en) | A kind of detection method of the intermediate for preparing Afatinib and its enantiomter | |
| CN114295764B (en) | Gel related substance detection method | |
| CN114609289B (en) | Method for detecting impurities in olmesartan medoxomil amlodipine compound preparation | |
| CN115792047B (en) | Method for detecting related substances of tedizolid phosphate intermediate | |
| CN114137133B (en) | Method for detecting related substances of naloxol-PEG derivative | |
| CN113960201B (en) | Method for detecting 4-dimethylaminopyridine in entecavir | |
| CN117250272A (en) | Detection method for simultaneously determining two genetic toxin impurities in propranolol hydrochloride |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20240319 Address after: No. 22, Yongwang Road, Biomedical Industry Base, Zhongguancun Science Park, Daxing District, Beijing 102600 Patentee after: Beijing Yunpeng Pengcheng Pharmaceutical Technology Co.,Ltd. Country or region after: Zhong Guo Patentee after: Yunpeng Pharmaceutical Group Co.,Ltd. Address before: No. 22, Yongwang Road, Biomedical Industry Base, Zhongguancun Science Park, Daxing District, Beijing 102600 Patentee before: Beijing Yunpeng Pengcheng Pharmaceutical Technology Co.,Ltd. Country or region before: Zhong Guo |
|
| TR01 | Transfer of patent right |