CN113640416A - Content determination method of levofloxacin hydrochloride tablets - Google Patents
Content determination method of levofloxacin hydrochloride tablets Download PDFInfo
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- CN113640416A CN113640416A CN202110923461.6A CN202110923461A CN113640416A CN 113640416 A CN113640416 A CN 113640416A CN 202110923461 A CN202110923461 A CN 202110923461A CN 113640416 A CN113640416 A CN 113640416A
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- levofloxacin hydrochloride
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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
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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
- G01N30/04—Preparation or injection of sample to be analysed
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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
- G01N30/34—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
- G01N30/74—Optical detectors
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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
- G01N30/86—Signal analysis
- G01N30/8624—Detection of slopes or peaks; baseline correction
- G01N30/8631—Peaks
- G01N30/8634—Peak quality criteria
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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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N2030/042—Standards
- G01N2030/047—Standards external
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Abstract
The invention discloses a content detection method of levofloxacin hydrochloride tablets, which adopts high performance liquid chromatography for detection, and the mobile phase is a mixture of the following components in volume ratio (1.5-2.0): 1 sodium hexane sulfonate solution and methanol. The invention provides a content detection method of levofloxacin hydrochloride tablets, which adopts sodium hexane sulfonate solution and methanol with a specific volume ratio as mobile phases and can effectively shorten retention time on the premise of ensuring that the chromatogram of the levofloxacin hydrochloride tablets has good peak shape and the separation degree and the theoretical plate number accord with the specification, thereby reducing the consumption of the mobile phases and improving the detection efficiency.
Description
Technical Field
The invention relates to the field of medicine quality control, in particular to a content detection method of levofloxacin hydrochloride tablets.
Background
Levofloxacin is a novel fluoroquinolone drug developed by first pharmaceutical corporation of japan in 1986, is an optically active L-isomer of ofloxacin, and is first marketed in japan at the end of 1993. Compared with the levofloxacin, the hydrochloride (levofloxacin hydrochloride) of the levofloxacin hydrochloride is relatively stable, has long storage time, relatively simple pharmaceutical process and easier absorption, and the medicament can quickly and widely permeate into various tissues and body fluids of a body. Can be used for treating infection caused by sensitive Staphylococcus, pneumococcus, Streptococcus pyogenes, hemolytic streptococcus, enterococcus, peptic streptococcus, gonococcus, Escherichia coli, Pseudomonas aeruginosa, Haemophilus influenzae, and Chlamydia trachomatis. The levofloxacin is completely absorbed by oral administration, so that the levofloxacin is beneficial to preparing an oral preparation which is convenient to use, and the medicines circulating in the market at present mainly comprise tablets and hard capsules.
The 2000 edition of Chinese pharmacopoeia (2004. supplement) adopts high performance liquid chromatography to measure the content of levofloxacin hydrochloride tablets, and adopts sodium hexane sulfonate solution-methanol (3:1) as a mobile phase. However, this method causes the retention time of levofloxacin hydrochloride and its main impurities to be too long, adversely affecting the working efficiency and increasing the consumption of mobile phase.
Therefore, under the condition of ensuring the accurate content measurement of the levofloxacin hydrochloride tablets, the retention time of the chromatogram is reduced as much as possible, and the method is very favorable for quality control in large-scale production of factories.
Disclosure of Invention
In view of this, the present invention provides a method for detecting the content of levofloxacin hydrochloride tablets, so as to reduce the retention time of chromatography.
The specific technical scheme is as follows:
the invention provides a content detection method of levofloxacin hydrochloride tablets, which adopts high performance liquid chromatography for detection, and the mobile phase is a mixture of the following components in volume ratio of (1.5-2.0): 1 sodium hexane sulfonate solution and methanol.
In some embodiments of the invention, the mobile phase is a mixture of 1.8:1 sodium hexane sulfonate solution and methanol.
In some embodiments of the invention, the chromatographic conditions comprise:
a chromatographic column: octadecylsilane chemically bonded silica;
the column temperature is 40 ℃;
the detection wavelength was 293 nm.
In some embodiments of the invention, the method comprises the steps of:
(1) dissolving levofloxacin hydrochloride tablets in a hydrochloric acid aqueous solution to obtain a sample stock solution;
(2) diluting the sample stock solution with the hydrochloric acid aqueous solution to obtain a sample test solution;
(3) injecting the test solution into a liquid chromatograph, recording the chromatogram, and reading the peak area S of levofloxacin hydrochlorideFor supplying to；
(4) According to the steps (1) to (3), measuring the levofloxacin contrast product by the same method, recording the chromatogram, and reading the peak area S of the levofloxacinTo pair；
(5) According to SFor supplying toAnd STo pairAnd determining the content of the levofloxacin hydrochloride tablets according to an external standard method.
In some embodiments of the invention, the concentration of the test solution is in the range of 0.06 to 0.14 mg/ml.
In some embodiments of the invention, the concentration of the test solution is in the range of 0.1 mg/ml.
In some embodiments of the invention, the concentration of the aqueous hydrochloric acid solution in steps (1) to (2) is 0.03 mol/ml.
Advantageous effects
The invention provides a content detection method of levofloxacin hydrochloride tablets, which adopts sodium hexane sulfonate solution and methanol with a specific volume ratio as mobile phases and can effectively shorten retention time on the premise of ensuring that the chromatogram of the levofloxacin hydrochloride tablets has good peak shape and the separation degree and the theoretical plate number accord with the specification, thereby reducing the consumption of the mobile phases and improving the detection efficiency.
Detailed Description
In order to make the objects, technical solutions and advantages of the present application more apparent, the technical solutions of the present application will be clearly and completely described below through specific embodiments.
In the following examples, those not indicated with specific conditions were performed according to conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
EXAMPLE 1 determination of the Mobile phase
The flow phase levofloxacin hydrochloride tablets of table 1 below were used for the detection, respectively.
TABLE 1
| Mobile phase 1 | Sodium hexane sulfonate solution-methanol (3:1) |
| Mobile phase 2 | Sodium hexane sulfonate solution-methanol (1.8:1) |
| Mobile phase 3 | Sodium hexane sulfonate solution-methanol (1.5:1) |
Wherein, the sodium hexane sulfonate solution is prepared as follows:
0.98g of sodium hexane sulfonate is taken and added with phosphate buffer (6.8 g of monopotassium phosphate is taken and dissolved by water and diluted to 1000ml, about 500ml of 0.05mol/L phosphoric acid is added, and the pH value is adjusted to 2.4.
The chromatographic conditions were as follows:
stationary phase: dima C18 column, 4.6X 200mm, 5um,
mobile phase: see Table 1
Flow rate: 1.0ml/min of the mixture is added,
detection wavelength: the wavelength of the light beam is 293nm,
column temperature: at a temperature of 40 c,
sample introduction amount: 20 μ l.
The detection steps are as follows:
(1) placing levofloxacin hydrochloride tablets (about equivalent to 50mg of levofloxacin) into a 50ml measuring flask, adding 0.03mol/ml hydrochloric acid solution to dissolve and dilute to scale, shaking up, and filtering;
(2) precisely measuring 5ml of the subsequent filtrate, placing the subsequent filtrate in a 50ml measuring flask, adding a mobile phase to dilute the subsequent filtrate to a scale, and shaking up to obtain a sample test solution;
(3) and (4) injecting 20 mu l of the test solution into a liquid chromatograph, and recording the chromatogram.
By adopting the mobile phase, the levofloxacin hydrochloride tablets have good peak shapes, the retention time of the main peak is relatively short when the ratio of the mobile phase 3 is (1.5:1), the retention time of the main peak is longer when the ratio of the mobile phase 1 (the mobile phase recorded in Chinese pharmacopoeia) is (3:1), the retention time of the mobile phase is about 10 minutes when the ratio of the mobile phase is (1.8:1) and the flow rate is 1.0ml/min, and the separation degree and the number of theoretical plates are both in accordance with the specification. Therefore, to reduce the consumption of the mobile phase, providing detection efficiency, the mobile phase according to the present invention is a mobile phase having a volume ratio of (1.5-2.0): 1-methanol, mobile phase 2 (1.8:1) is preferably used as the content determination mobile phase for levofloxacin hydrochloride tablets according to the invention.
EXAMPLE 2 determination of the Linear Range of the assay
Weighing appropriate amount of levofloxacin reference, precisely weighing, adding mobile phase for dissolving, and quantitatively diluting to obtain a stock solution containing levofloxacin reference 0.2 mg/ml; precisely measuring 3ml, 4ml, 5ml, 6ml and 7ml, respectively placing into 10ml measuring flask, adding mobile phase to dilute to scale, shaking, precisely measuring 20 μ l of sample solution, injecting into liquid chromatograph, recording chromatogram, and referring to example 1 (mobile phase selected mobile phase 2). The test results are given in the following table: the results are shown in Table 1 below:
TABLE 1 measurement results of the Linear relationship test
The regression analysis confirms that the sample injection concentration is in a good linear relation within the range of 0.06mg/ml to 0.14mg/ml, and can meet the requirement of content determination.
Example 3 content measurement of levofloxacin hydrochloride tablets
The content of the self-made levofloxacin hydrochloride tablets of 3 batches is detected by adopting the high performance liquid chromatography detection method provided by the invention.
The chromatographic conditions were as follows:
stationary phase: dima C18 column, 4.6X 200mm, 5um,
mobile phase: sodium hexane sulfonate solution-methanol (1.8:1),
flow rate: 1.0ml/min of the mixture is added,
detection wavelength: the wavelength of the light beam is 293nm,
column temperature: at a temperature of 40 c,
sample introduction amount: 20 μ l.
The detection steps are as follows:
(1) placing levofloxacin hydrochloride tablets (about equivalent to 50mg of levofloxacin) into a 50ml measuring flask, adding 0.03mol/ml hydrochloric acid solution to dissolve and dilute to scale, shaking up, and filtering;
(2) precisely measuring 5ml of the subsequent filtrate, placing the subsequent filtrate in a 50ml measuring flask, adding a mobile phase to dilute the subsequent filtrate to a scale, and shaking up to obtain a sample test solution;
(3) injecting 20 μ l of the sample solution into a liquid chromatograph, recording chromatogram, and reading the peak area S of levofloxacin hydrochlorideFor supplying to；
(4) Taking levofloxacin reference (130455-To pair；
(5) According to SFor supplying toAnd STo pairThe content of levofloxacin hydrochloride tablets was determined by external standard method, and the results are shown in table 2 below.
TABLE 2 results of content measurement
Note: the contents in table 2 refer to contents in relative amounts.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention.
Claims (7)
1. A content detection method of levofloxacin hydrochloride tablets adopts high performance liquid chromatography for detection, and is characterized in that a mobile phase is a mixture of levofloxacin hydrochloride tablets and levofloxacin hydrochloride tablets in a volume ratio of (1.5-2.0): 1 sodium hexane sulfonate solution and methanol.
2. The method for detecting the content of levofloxacin hydrochloride tablets according to claim 1, wherein the mobile phase is a mixture of the mobile phase and the levofloxacin hydrochloride tablet at a volume ratio of 1.8:1 sodium hexane sulfonate solution and methanol.
3. The method for detecting the content of levofloxacin hydrochloride tablets according to claim 1, wherein the chromatographic conditions comprise:
a chromatographic column: octadecylsilane chemically bonded silica;
the column temperature is 40 ℃;
the detection wavelength was 293 nm.
4. The method for detecting the content of levofloxacin hydrochloride tablets according to claim 1, wherein the method comprises the following steps:
(1) dissolving levofloxacin hydrochloride tablets in a hydrochloric acid aqueous solution to obtain a sample stock solution;
(2) diluting the sample stock solution with the hydrochloric acid aqueous solution to obtain a sample test solution;
(3) injecting the sample solution into liquid chromatograph, and recording chromatogramDrawing, and reading the peak area S of levofloxacin hydrochlorideFor supplying to；
(4) According to the steps (1) to (3), measuring the levofloxacin contrast product by the same method, recording the chromatogram, and reading the peak area S of the levofloxacinTo pair；
(5) According to SFor supplying toAnd STo pairAnd determining the content of the levofloxacin hydrochloride tablets according to an external standard method.
5. The method for detecting the content of levofloxacin hydrochloride tablets according to claim 1, wherein the concentration of the test solution is in the range of 0.06-0.14 mg/ml.
6. The method for detecting the content of levofloxacin hydrochloride tablets according to claim 5, wherein the concentration of the test solution is in the range of 0.1 mg/ml.
7. The method for detecting the content of levofloxacin hydrochloride tablets according to claim 4, wherein the concentration of the aqueous hydrochloric acid solution in the steps (1) to (2) is 0.03 mol/ml.
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2021
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