CN104965031B - Content measuring method for compound ketoprofen and omeprazole sustained-release capsules - Google Patents

Abstract

The invention discloses a content measuring method for compound ketoprofen and omeprazole sustained-release capsules. The content measuring method also can be used for detecting other solution containing ketoprofen and omeprazole such as releasing rate solution of the compound ketoprofen and omeprazole sustained-release capsules. Octadecylsilane chemically bonded silica is used as a filler of a chromatographic column; a mixed solvent of acetonitrile and 0.02 mol/l potassium dihydrogen phosphate [with a pH (potential of hydrogen) value which is adjusted by phosphoric acid solution until the pH value reaches 4.0] is used as mobile phase, and the volume ratio of the acetonitrile to the 0.02 mol/l potassium dihydrogen phosphate is 45:55; detecting wavelengths are 210-240 nanometers and 275-285 nanometers. The content measuring method particularly includes respectively preparing reference solution of ketoprofen and reference solution of the omeprazole; preparing test solution of the compound ketoprofen and omeprazole sustained-release capsules; precisely measuring 10-20 micro-liters of the test solution and 10-20 micro-liters of the reference solution, injecting the measured test solution and the measured reference solution in a liquid chromatograph, and recording chromatogram maps; respectively computing the contents of the ketoprofen and the omeprazole in samples by the aid of peak areas according to an external standard process.

Description

Content Determination Method of Compound Ketoprofen Omeprazole Sustained Release Capsules

technical field

The invention belongs to the field of drug analysis and detection, in particular to a method for determining the content and release degree of compound ketoprofen omeprazole sustained-release capsules.

Background technique

Compound Ketoprofen Omeprazole Sustained-release Capsules is the first compound preparation composed of Ketoprofen and Omeprazole, developed by Sweden Meda Pharmaceutical Company, with a trade name of Axorid, which is effective in alleviating osteoarthritis, rheumatoid arthritis When treating the pain of diseases such as inflammation and ankylosing spondylitis, it can effectively prevent and treat adverse reactions, effectively prevent gastric ulcer and bleeding, and double protect the lives and health of patients. In the prior art, there is no disclosure about a method for simultaneously measuring the content of two medicinal components, ketoprofen and omeprazole, in a solution. The content detection method of compound ketoprofen omeprazole sustained-release capsule of the present invention, use a kind of mobile phase condition and can determine the content of two kinds of components in this compound preparation simultaneously; The advantage of this detection method is: easy and simple to operate; Measurement result Accurate and reliable; strong specificity; short detection time.

Contents of the invention

The purpose of the present invention is to provide a kind of HPLC assay method that can conveniently and rapidly detect the content of compound ketoprofen omeprazole slow-release capsule and the release amount in the release solution.

In order to achieve the above object, the following technical solutions adopted by the present invention are realized.

A method for detecting the content of compound ketoprofen omeprazole sustained-release capsules, characterized in that:

(1) Chromatographic conditions:

Stationary phase: choose octadecylsilane bonded silica gel as the chromatographic column;

Mobile phase: a mixed solvent composed of component A and component B with a volume ratio of 35:65 to 55:45 is used as the mobile phase; the component A is acetonitrile, and the concentration of component B is 0.01 to 0.05mol/L Potassium dihydrogen phosphate solution, sodium dihydrogen phosphate solution or ammonium dihydrogen phosphate solution, adjust the pH value to 2.5-4.5 with phosphoric acid solution;

Preferably: the mixed solvent of acetonitrile-0.02mol/l potassium dihydrogen phosphate solution with a volume ratio of 45:55 is used as mobile phase, and the pH value is adjusted to 4.0 with phosphoric acid solution;

Detection wavelength: 210-240nm and 275-285nm; preferably 279nm;

Column temperature: 25-35°C; preferably 30°C;

Flow rate: flow rate 0.8 ~ 1.2ml/min; preferably 1.0ml/min.

(2) Experimental materials

a. Preparation of reference substance solution: Accurately weigh about 200 mg of ketoprofen reference substance, put it into a 100ml brown measuring bottle, add 50% acetonitrile solution to dissolve and dilute to the mark, shake well, as ketoprofen stock solution; Omeprazole reference substance is about 20mg, add 20ml of ethanol and about 60ml of phosphate buffer (pH11.0), shake to dissolve, dilute to the mark with phosphate buffer (pH11.0), shake well, and use as Omeprazole Prazole stock solution. Precisely measure 10, 15, 20ml of ketoprofen stock solution and 10ml of omeprazole stock solution respectively, put them into a 100ml measuring bottle, dilute to the mark with water, and serve as the comparison of 100mg/20mg, 150mg/20mg and 200mg/20mg specifications respectively product solution.

B. the preparation of test solution: get 10 compound ketoprofen omeprazole slow-release capsules, get the content and put it into a 500ml measuring bottle, add phosphate buffer (pH11.0) 50ml, 1mol/L sodium hydroxide Solution 10ml and ethanol 100ml, shake to dissolve the pellets, add phosphate buffer (pH11.0) to dilute to the mark, shake well, centrifuge, accurately measure 5ml of the supernatant, put it into a 100ml measuring bottle, add water to dilute to the mark, shake Get it evenly.

c. Preparation of phosphate buffer (pH11.0): Take 110ml of 0.25mol/L sodium phosphate solution and 220ml of 0.5mol/L disodium hydrogen phosphate solution, dilute with water to 1000ml, and shake well.

(3) Accurately draw respectively 10-20 μl of the reference substance solution and the test solution, preferably 10 μl, inject into the liquid chromatograph, measure and calculate the content of ketoprofen and omeprazole.

The present invention has the advantages of simultaneously measuring the content of ketoprofen and omeprazole in compound ketoprofen omeprazole sustained-release capsules through optimized high performance liquid chromatography, the method is easy to operate, and the measurement result is accurate and reliable , good stability, good repeatability, strong specificity, and short detection time.

Description of drawings

The HPLC collection of illustrative plates of Fig. 1 reference substance (the retention times of omeprazole and ketoprofen are respectively 5.1min and 11.8min.)

The HPLC pattern of figure 2 negative control solution

detailed description

The content of the present invention is explained through the following specific embodiments, but the present invention is not limited. Because the compound ketoprofen omeprazole sustained-release capsule has 3 specifications, only the 100mg/20mg specification is proposed as an example for illustration.

Instruments and reagents:

Instrument: High performance liquid chromatography: SHIMADZU LC-20AT SPD-M20A; Chromatographic column: R&C (Research and Creation) C ₁₈ column (250mm×4.6mm, 5μm).

Reference substance: Ketoprofen (National Institute for the Control of Pharmaceutical and Biological Products 100337-201104); Omeprazole (National Institute for the Control of Pharmaceutical and Biological Products 100367-201305).

Test product: Compound Ketoprofen Omeprazole Sustained Release Capsules (Shandong Academy of Pharmaceutical Sciences).

Reagents: chromatographic grade acetonitrile (Dikmapure Company), potassium dihydrogen phosphate, sodium dihydrogen phosphate, ammonium dihydrogen phosphate (Sinopharm Group), phosphoric acid (analytical grade).

Solution preparation:

Preparation of reference substance solution: Accurately weigh about 200 mg of ketoprofen reference substance, put it into a 100ml brown measuring bottle, add 50% acetonitrile solution to dissolve and dilute to the mark, shake well, and use it as ketoprofen stock solution; accurately weigh Ogilvy About 20mg of the prazole reference substance, add 20ml of ethanol and about 60ml of phosphate buffer (pH11.0), shake to dissolve, dilute to the mark with phosphate buffer (pH11.0), shake well, and use as omeprazole stock solution. Accurately measure 10ml of ketoprofen stock solution and 10ml of omeprazole stock solution respectively, put them into a 100ml measuring bottle, add water to dilute to the mark, shake well, and obtain.

The preparation of need testing solution: get compound ketoprofen omeprazole slow-release capsule (specification 100mg/20mg) 10, get content and put into 500ml measuring bottle, add phosphate buffer saline (pH11.0) 50ml, 1mol/ 10ml of sodium hydroxide solution and 100ml of ethanol, shake to dissolve the pellets, add phosphate buffer (pH11.0) to dilute to the mark, shake well, centrifuge at 9000r/min for 15min, accurately measure 5ml of the supernatant and put it into a 100ml measuring bottle , add water to dilute to the mark, shake well, that is.

The following is the screening experiment of the chromatographic conditions and the solvent of the content determination method of the compound ketoprofen omeprazole sustained-release capsules of the present invention.

1. Selection of mobile phase

Referring to the relevant literature and combined with the specific conditions of the test, acetonitrile-phosphate buffer (0.01mol/L sodium dihydrogen phosphate, adjusted to pH 4.0 with phosphoric acid) (55:45, v/v), acetonitrile-phosphate buffer was successively selected. (0.02mol/L ammonium dihydrogen phosphate, adjust the pH value to 3.5 with phosphoric acid) (45:55, v/v), acetonitrile-phosphate buffer (0.03mol/L potassium dihydrogen phosphate, adjust the pH value to 3.0 with phosphoric acid ) (35:65, v/v), acetonitrile-phosphate buffer (0.02mol/L potassium dihydrogen phosphate, adjusted to pH 4.0 with phosphoric acid) (45:55, v/v) is the mobile phase, each mobile phase The separation effects under the conditions are shown in Table 1.

Table 1 mobile phase selection results

2. Selection of flow rate

Select the flow rate as 0.8, 1.0 and 1.2ml/min to investigate, the difference is small, and finally determine the flow rate as 1.0ml/min.

3. Selection of column temperature

The column temperature was selected as 25, 30 and 35°C for investigation, the separation became worse at 35°C, and finally the column temperature was determined to be 30°C.

4. Selection of solvent

Select acetonitrile, 50% acetonitrile, phosphate buffer (pH11.0) successively, first add 50ml of phosphate buffer (pH11.0), 10ml of 1mol/L sodium hydroxide solution and 100ml of ethanol to dissolve the pellets, then add Phosphate buffer (pH11.0) to the mark is the solvent. The solvent selection results are shown in Table 2.

Table 2 Solvent selection results

The following is an experiment to verify the assay effect after the chromatographic conditions of the content assay method of the present invention are determined.

1. Chromatographic conditions

A chromatographic column filled with octadecylsilane-bonded silica gel, with acetonitrile-phosphate buffer (0.02mol/L potassium dihydrogen phosphate, adjusted to pH 4.0 with phosphoric acid) (45:55, v/v) as the flow phase, the column temperature is 30°C, the flow rate is 1.0ml/min, and the detection wavelength is 279nm.

2. Exclusive research

Mix the excipients evenly according to the proportion of the prescription, weigh an appropriate amount and put it into a 500ml measuring bottle, prepare according to the preparation method of the test solution to obtain a negative reference solution, accurately measure 10 μl and inject it into the liquid chromatograph, and record the chromatogram. The results are shown in accompanying drawing 2, as can be seen from Figure 2, the auxiliary materials of this product do not affect the chromatographic peaks of the two main components.

3. Linearity test and range

Accurately weigh an appropriate amount of ketoprofen and omeprazole reference substance, add diluent to dissolve and dilute to make a solution containing about 1000 μg of ketoprofen and 200 μg of omeprazole in every 1 ml, as a stock solution. Accurately measure the appropriate amount of stock solution, and make each 1ml of ketoprofen/omeprazole 70μg/14μg, 80μg/16μg, 90μg/18μg, 100μg/20μg, 110μg/22μg, 120μg/24μg, 130μg/26μg as a linear solution ; Precisely measure 10 μl of the linear solution and inject it into the chromatograph, and record the chromatogram. Take peak area A as ordinate, concentration (C) as abscissa, carry out linear regression, get ketoprofen regression equation: A=1.445×10 ⁴ C+0.361×10 ⁴ , linear correlation coefficient r=0.9997, Omega Azole regression equation: A=1.552×10 ⁴ C+367.1, linear correlation coefficient r=0.9993; among them, ketoprofen has a good linear relationship with its peak area in the range of 69.96-129.92 μg/ml, and omeprazole has a good linear relationship between 14.06-26.11 It has a good linear relationship with its peak area in the range of μg/ml.

4. Stability test

Precisely draw 10 μ l of the test solution, inject it into the liquid chromatograph at 0, 3, 6, 9, and 12 hours respectively, record the peak area, calculate the peak area of ketoprofen and omeprazole, and the RSD is 0.53%, 0.61% respectively , indicating that the test solution is stable within 12h.

5. Precision test

Precisely draw 10 μl of the reference substance solution, repeat the injection 6 times, record the peak area, calculate the peak area of ketoprofen and omeprazole, and the RSDs are 0.13% and 0.21%, respectively.

6. Repeatability test

Accurately draw need testing solution 10 μ l, parallel 6 parts, measure the content of every part of need testing according to optimal chromatographic conditions, inject liquid chromatograph, record peak area, and every capsule contains ketoprofen and omeprazole respectively on average 99.54% (RSD=0.38%), 99.43% (RSD=0.43%) of the labeled amount.

7. Recovery test

According to the prescription ratio, accurately measure 3 parts of ketoprofen stock solution and omeprazole stock solution 8,10,12ml, add auxiliary materials, put into 100ml volumetric flask, measure according to the above-mentioned preferred chromatographic conditions, and calculate the recovery rate. The average recoveries of ketoprofen and omeprazole were 98.83% (RSD=0.46%) and 99.99% (RSD=0.76%), respectively.

8. Content Determination of Compound Ketoprofen Omeprazole Sustained Release Capsules

Take 10 compound ketoprofen omeprazole sustained-release capsules (batch numbers FFTA01, FFTA02, FFTA03), put the contents into a 500ml measuring bottle, add 50ml of phosphate buffer (pH11.0), 1mol/L sodium hydroxide solution 10ml and 100ml ethanol, shake to dissolve the pellets, add phosphate buffer (pH11.0) to dilute to the mark, shake well, centrifuge at 9000r/min for 15min, accurately measure 5ml of the supernatant, put it into a 100ml measuring bottle, and dilute with water to scale, shake well, and determine with high performance liquid chromatography.

a. A chromatographic column with octadecylsilane bonded silica gel as a filler, with acetonitrile-phosphate buffer (0.02mol/L potassium dihydrogen phosphate, adjust the pH value to 4.0 with phosphoric acid) (45:55, v/v) As the mobile phase, the column temperature is 30°C, the flow rate is 1.0ml/min, and the detection wavelength is 279nm.

b. Accurately draw respectively 10 μ l of the reference substance solution and the test solution under the content determination item, inject the liquid chromatograph, measure, and calculate the content of ketoprofen and omeprazole, the results are shown in Table 3.

The content determination result of ketoprofen and omeprazole in the compound ketoprofen omeprazole sustained-release capsule of table 3

9 Determination of release rate of compound ketoprofen omeprazole sustained-release capsules

Get the release solution of compound ketoprofen omeprazole sustained-release capsules (batch numbers FFTA01, FFTA02, FFTA03) at 6h, filter, get the continued filtrate, and measure with high performance liquid chromatography:

a. A chromatographic column with octadecylsilane bonded silica gel as a filler, with acetonitrile-phosphate buffer (0.02mol/L potassium dihydrogen phosphate, adjust the pH value to 4.0 with phosphoric acid) (45:55, v/v) As the mobile phase, the column temperature is 30°C, the flow rate is 1.0ml/min, and the detection wavelength is 279nm.

b. Accurately draw respectively 10 μ l of the reference substance solution and the test solution under the release rate item, inject into the liquid chromatograph, measure and calculate the release amount of ketoprofen and omeprazole, the results are shown in Table 4.

The 6h release measurement result of table 4 compound ketoprofen omeprazole sustained-release capsules

Claims (5)

1. a method for assaying compound ketoprofen omeprazole sustained-release capsule, is characterized in that: adopt high performance liquid chromatography (HPLC) to detect, and detection condition is as follows: Stationary phase: choose octadecylsilane bonded silica gel as the chromatographic column; Mobile phase: a mixed solvent composed of component A and component B with a volume ratio of 35:65 to 55:45 is used as the mobile phase; the component A is acetonitrile; the concentration of component B is 0.01 to 0.05mol/L Potassium dihydrogen phosphate solution, sodium dihydrogen phosphate solution or ammonium dihydrogen phosphate solution, adjust the pH value to 2.5-4.5 with phosphoric acid solution; Detection wavelength: 279nm; Column temperature: 25～35℃; Flow rate: flow rate 0.8 ~ 1.2ml/min; Experimental Materials: a. Preparation of reference substance solution: Accurately weigh about 200 mg of ketoprofen reference substance, put it into a 100ml brown measuring bottle, add 50% acetonitrile solution to dissolve and dilute to the mark, shake well, as ketoprofen stock solution; Omeprazole reference substance is about 20mg, add ethanol 20ml and pH 11.0 phosphate buffer about 60ml, shake to dissolve, dilute to the mark with pH 11.0 phosphate buffer, shake well, as omeprazole Stock solution: Accurately measure 10, 15, 20ml of ketoprofen stock solution and 10ml of omeprazole stock solution respectively, put them into a 100ml measuring bottle, dilute to the mark with water, and use them as 100mg/20mg, 150mg/20mg and 200mg/20mg respectively Standard reference substance solution; B. The preparation of test solution: get 10 compound ketoprofen omeprazole slow-release capsules, get the content and put it into a 500ml measuring bottle, add phosphate buffer 50ml of pH11.0, 1mol/L sodium hydroxide solution 10ml and 100ml ethanol, shake to dissolve the pellets, add pH 11.0 phosphate buffer solution to dilute to the mark, shake well, centrifuge, accurately measure 5ml of the supernatant, put it into a 100ml measuring bottle, add water to dilute to the mark, and shake well; c. Preparation of pH 11.0 phosphate buffer: take 110ml of 0.25mol/L sodium phosphate solution and 220ml of 0.5mol/L disodium hydrogen phosphate solution, dilute with water to 1000ml, and shake well; Accurately draw 10-20 μl each of the reference substance solution and the test solution, inject it into a liquid chromatograph, measure and calculate the contents of ketoprofen and omeprazole. 2. the assay method of a kind of compound ketoprofen omeprazole slow-release capsule according to claim 1, is characterized in that: component A described in high performance liquid chromatography detection condition is acetonitrile, component B It is a potassium dihydrogen phosphate solution with a concentration of 0.02mol/L, and the pH value is adjusted to 4.0 with a phosphoric acid solution, and the volume ratio is 45:55. 3. the assay method of a kind of compound ketoprofen omeprazole slow-release capsule according to claim 1, is characterized in that: the flow velocity described in high performance liquid chromatography detection condition is 1.0ml/min, column temperature is 30°C. 4. the content assay method of a kind of compound ketoprofen omeprazole sustained-release capsule according to claim 1, is characterized in that: the sample size of high performance liquid chromatography detection condition is 10 μ l. 5. the assay method of a kind of compound ketoprofen omeprazole sustained-release capsule according to claim 1, is characterized in that: this method is detecting the content of compound ketoprofen omeprazole sustained-release capsule, and The application of solution detection of release rate of compound ketoprofen omeprazole sustained-release capsules.