CN110542737B - Detection method for dissolution determination of flunarizine hydrochloride capsule - Google Patents
Detection method for dissolution determination of flunarizine hydrochloride capsule Download PDFInfo
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Abstract
The invention discloses a detection method for dissolution determination of flunarizine hydrochloride capsules, which comprises the following steps: the dissolution method is a paddle method, 100 revolutions per minute, and a medium added with sodium dodecyl sulfate is a dissolution medium; detecting the eluate by high performance liquid chromatography, and using octadecylsilane chemically bonded silica as filler; an ultraviolet detector with the detection wavelength of 251-255 nm; the column temperature is 25-45 ℃; the flow rate is 0.9-1.1 ml/min; the mobile phase is methanol and phosphate buffer. The method solves the problem of filter membrane adsorption of flunarizine hydrochloride capsules, the obtained chromatographic peak has good shape and high column efficiency, the obtained result is more accurate, the service life of the chromatographic column is prolonged, and the test cost is reduced.
Description
Technical Field
The invention belongs to the technical field of analytical chemistry, and particularly relates to a detection method for dissolution determination of flunarizine hydrochloride capsules.
Background
The flunarizine hydrochloride capsule is a vasodilatation medicine and is mainly used for preventing and treating typical or atypical migraine and symptomatic treatment of vertigo caused by vestibular dysfunction.
The flunarizine hydrochloride is a selective calcium antagonist, can block excessive calcium ions from entering cells through a membrane, and prevents the excessive calcium load in the cells. It also has effects in preventing large amount of calcium from entering neuron during ischemia and anoxia, improving brain microcirculation and neuron metabolism, inhibiting cerebral vasospasm, platelet aggregation and blood viscosity increase, and stabilizing cell pleural membrane. The product has high lipid solubility, and is easy to penetrate blood brain barrier. It has no influence on cardiac contraction and conduction.
At present, the detection method of flunarizine hydrochloride capsules is recorded in the Chinese pharmacopoeia 2015 edition and includes a detection method for flunarizine hydrochloride dissolution measurement. The dissolution method comprises taking 1 granule of the product, taking 600ml hydrochloric acid solution (diluted hydrochloric acid 24ml and water to 1000ml) as dissolution medium according to dissolution and release rate determination method (general rule 0931 first method), rotating at 100 rpm, taking 10ml solution after 30min, filtering, and taking the subsequent filtrate as sample solution. Adopting high performance liquid chromatography, using octadecylsilane chemically bonded silica as filler, methanol-phosphate buffer (taking 1.36g of potassium dihydrogen phosphate, adding water to dissolve and dilute into 1000ml, adding 4ml of triethylamine, adjusting pH value to 3.5 with phosphoric acid) (75:25) as mobile phase; the detection wavelength is 253nm, and the column temperature is 35 ℃. Wherein the dissolution rate is required to be hydrochloric acid medium (dilute hydrochloric acid 5.6ml → 1L).
In the research on the dissolution curve of the flunarizine hydrochloride capsule, the examination on the dissolution curves of other media is carried out, and the filter membrane adsorption phenomenon exists in the other media, and the filter membrane adsorption phenomenon does not exist after the sodium dodecyl sulfate is added. The method mainly inspects SDS medium with pH of 6.8+ 0.4%, and researches and comparisons show that the detection method recorded in Chinese pharmacopoeia 2015 edition has better effects on both chromatographic column peak type and column efficiency when the dissolution of flunarizine hydrochloride capsule pharmacopoeia medium (hydrochloric acid medium) is measured. However, in the study of the dissolution curve of SDS medium with pH6.8+ 0.4%, potassium dihydrogen phosphate, which is used as the mobile phase of the salt, reacts with sodium dodecyl sulfate in the sample solvent, so that the chromatographic peak is branched, the column performance is lowered, and the life of the chromatographic column is shortened. The method is not suitable for the research of the dissolution of the flunarizine hydrochloride capsule no longer in consideration of result accuracy or economy by combining the influence of filter membrane adsorption.
Therefore, a new method is urgently needed to be developed for the research of the dissolution of the flunarizine hydrochloride capsule, and the method not only aims at the dissolution measurement of the hydrochloric acid medium of the flunarizine hydrochloride capsule, but also has a better effect on the dissolution measurement of other added surfactants.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: provides a detection method for dissolution determination of flunarizine hydrochloride capsules, which aims to solve the problems in the prior art.
The technical scheme adopted by the invention is as follows: a detection method for dissolution determination of flunarizine hydrochloride capsules comprises the following steps: the dissolution method is a paddle method, 100 revolutions per minute, and a medium added with sodium dodecyl sulfate is a dissolution medium; detecting the eluate by high performance liquid chromatography, and using octadecylsilane chemically bonded silica as filler; an ultraviolet detector with the detection wavelength of 251-255 nm; the column temperature is 25-45 ℃; the flow rate is 0.9-1.1 ml/min; the mobile phase is methanol and phosphate buffer.
Sodium dodecyl sulfate is added in an amount of 0.01 to 1.0% (w/V), preferably 0.4% (w/V).
The ratio of mobile phase methanol to phosphate buffer was 75: 25.
The phosphate buffer solution is sodium dihydrogen phosphate solution, and the concentration of phosphate is 0.01 mol/L.
4ml triethylamine were added per 1000m phosphate buffer and the pH was adjusted to 3.5. + -. 0.05% with phosphoric acid.
The detection wavelength adopted by the invention is 251-255nm, because the maximum absorption wavelength of the flunarizine hydrochloride is 253nm, and the content and dissolution of the flunarizine hydrochloride are only used for detecting the quantity of the flunarizine, and the preferred detection wavelength is 253 nm.
The column temperature adopted by the invention is 25-45 ℃ according to the conventional chromatographic column use temperature and the principle of convenient temperature control. To avoid precipitation of sodium lauryl sulfate due to low temperature, 35 ℃ is more preferable.
The flow rate setting mentioned in the present invention is common knowledge of the skilled person, and the common range is generally 0.5ml/min to 2ml/min, and in order to obtain better separation effect and improve test efficiency, the flow rate is preferably 0.9-1.1ml/min, more preferably 1.0 ml/min.
The invention has the beneficial effects that: compared with the prior art, the invention has the following effects:
(1) the method of the invention adopts the medium added with the lauryl sodium sulfate as the dissolution medium to solve the problem of filter membrane adsorption of the flunarizine hydrochloride capsule.
(2) The chromatographic peak obtained by the method has good peak shape and high column efficiency, and the obtained result is more accurate. And the service life of the chromatographic column is prolonged, and the test cost is reduced.
(3) The phosphate in the mobile phase adopted by the invention is sodium dihydrogen phosphate, the solubility of the sodium dihydrogen phosphate is higher, the damage to a chromatographic column is smaller, and the reagents of the adopted mobile phase are common reagents and are cheap and easy to obtain;
(4) the method has simple operation process, good accuracy and repeatability;
(5) the method of the invention adopts instruments and equipment which are conventional articles, the test parameters are also conventional parameters, no harsh conditions are generated, the cost is low, and the conditions of most laboratories can meet the requirements.
Drawings
FIG. 1 is a blank chromatogram obtained by dissolving out Flunarizine hydrochloride capsule pH6.8+ 0.4% SDS
FIG. 2 is a control chromatogram obtained by dissolving out Flunarizine hydrochloride capsule pH6.8+ 0.4% SDS
FIG. 3 is a chromatogram of a test sample obtained by dissolving flunarizine hydrochloride capsule pH6.8+ 0.4% SDS
FIG. 4 is a chromatogram of a sample obtained by dissolving potassium dihydrogen phosphate in SDS at pH6.8+ 0.4% as a mobile phase.
Detailed Description
The invention is further described with reference to the accompanying drawings and specific embodiments.
As shown in fig. 1-3, a method for detecting dissolution measurement of flunarizine hydrochloride capsules, the dissolution method is a paddle method, 100 rpm, and a medium added with sodium dodecyl sulfate is a dissolution medium; detecting the eluate by high performance liquid chromatography, and using octadecylsilane chemically bonded silica as filler; an ultraviolet detector with the detection wavelength of 251-255 nm; the column temperature is 25-45 ℃; the flow rate is 0.9-1.1 ml/min; the mobile phase is methanol and phosphate buffer.
Sodium dodecyl sulfate is added in an amount of 0.01 to 1.0% (w/V), preferably 0.4% (w/V).
The ratio of mobile phase methanol to phosphate buffer was 75: 25.
The phosphate buffer solution is sodium dihydrogen phosphate solution, and the concentration of phosphate is 0.01 mol/L.
4ml triethylamine was added per 1000ml phosphate buffer and the pH was adjusted to 3.5. + -. 0.05 with phosphoric acid.
The phosphate in the mobile phase adopted by the invention is sodium dihydrogen phosphate, the solubility of the sodium dihydrogen phosphate is higher, and the damage to the chromatographic column is smaller.
The detection wavelength adopted by the invention is 251-255nm, because the maximum absorption wavelength of the flunarizine hydrochloride is 253nm, and the content and dissolution of the flunarizine hydrochloride are only used for detecting the quantity of the flunarizine, and the preferred detection wavelength is 253 nm.
The column temperature adopted by the invention is 25-45 ℃ according to the conventional chromatographic column use temperature and the principle of convenient temperature control. To avoid precipitation of sodium lauryl sulfate due to low temperature, 35 ℃ is more preferable.
The flow rate setting mentioned in the present invention is common knowledge of the skilled person, and the common range is generally 0.5ml/min to 2ml/min, and in order to obtain better separation effect and improve test efficiency, the flow rate is preferably 0.9-1.1ml/min, more preferably 1.0 ml/min.
The dissolution determination method of the flunarizine hydrochloride capsule comprises the following steps:
preparing a reference substance solution: precisely weighing a proper amount of flunarizine hydrochloride reference substance, quantitatively diluting with dissolution medium to prepare a solution containing 8.3 mu g of flunarizine per 1ml, and shaking up to obtain the reference substance solution.
Preparing a test solution: taking one capsule, placing into a dissolution cup filled with 600ml dissolution medium, and collecting 1ml filtrate after 30min as test solution.
Precisely measuring 20 μ l of each of the reference solution and the sample solution, injecting into a liquid chromatograph, and recording chromatogram. Calculating according to the peak area by an external standard method to obtain the content.
Example 1: detection method for dissolution determination of flunarizine hydrochloride capsule
Instruments and conditions:
high performance liquid chromatograph: DIONEXU 3000;
a chromatographic column: YMC-PackODSC18(4.6 × 250mm, 5 μm);
mobile phase: methanol-phosphate buffer (taking 1.20g of sodium dihydrogen phosphate, adding water to dissolve and dilute into 1000ml, adding 4ml of triethylamine, and adjusting the pH value to 3.5 by using phosphoric acid) (75: 25);
column temperature: 35 ℃;
flow rate: 1.0 ml/min;
detection wavelength: 253 nm;
sample introduction volume: 20 μ l.
The implementation steps are as follows:
(1) preparation of dissolution medium: SDS2.00g, sodium dihydrogen phosphate 6.00g (7.80 g sodium dihydrogen phosphate dihydrate) and sodium hydroxide 0.90g were weighed and diluted to 1000ml with water.
(2) Placing flunarizine hydrochloride 10.89mg and 11.02mg in a 100ml measuring flask, adding 10ml of ethanol to dissolve, adding a dissolving medium to dilute to scale, and shaking up. Precisely measuring 5ml, placing into a 50ml measuring flask, adding dissolution medium, diluting to scale, and shaking to obtain reference solution.
(3) Taking one capsule, placing into a dissolution cup filled with 600ml dissolution medium, and collecting 1ml filtrate after 30min as test solution.
(4) Precisely measuring 20 μ l of each of the reference solution and the sample solution, injecting into a liquid chromatograph, and recording chromatogram. A blank solvent control test was also performed. The content was calculated as peak area by external standard method. The dissolution rate is more than 90% in 30 min. The results are shown in FIGS. 1-3.
Example 2: a detection method for dissolution determination of flunarizine hydrochloride capsules comprises the following steps:
instruments and conditions:
high performance liquid chromatograph: DIONEXU 3000;
a chromatographic column: YMC-PackODSC18(4.6 × 250mm, 5 μm);
mobile phase: methanol-phosphate buffer (taking 1.20g of sodium dihydrogen phosphate, adding water to dissolve and dilute into 1000ml, adding 4ml of triethylamine, and adjusting the pH value to 3.5 by using phosphoric acid) (75: 25);
column temperature: 35 ℃;
flow rate: 1.0 ml/min;
detection wavelength: 253 nm;
sample introduction volume: 20 μ l.
The implementation steps are as follows:
(1) preparation of dissolution medium (adding SDS): SDS2.00g, sodium dihydrogen phosphate 6.00g (7.80 g sodium dihydrogen phosphate dihydrate) and sodium hydroxide 0.90g were weighed and diluted to 1000ml with water.
(2) Preparation of dissolution medium (without SDS): 6.00g of sodium dihydrogen phosphate and 0.90g of sodium hydroxide were weighed and diluted with water to 1000 ml.
(3) Taking one capsule, placing into a dissolution cup filled with 600ml dissolution medium, discarding 1ml, 2ml, 3ml, 5ml, 8ml and 10ml respectively after 30min, and using as filter membrane to adsorb the sample solution. And meanwhile, centrifuging the solution, and taking the supernatant after centrifugation as a centrifuged solution.
(4) Precisely measuring 20 μ l of each of the reference solution and the sample solution, injecting into a liquid chromatograph, and recording chromatogram. A blank solvent control test was also performed. The content was calculated as peak area by external standard method.
The results of the effect of the addition of the surfactant on the adsorption on the filter are shown in table 1.
TABLE 1 adsorption results of different dissolution media filters
This makes it possible to substantially eliminate the influence of adsorption on the filter membrane by adding the surfactant.
Example 3: detection method for dissolution determination of flunarizine hydrochloride capsule in prior art(control case)
Instruments and conditions:
high performance liquid chromatograph: DIONEX U3000;
a chromatographic column: YMC-Pack ODS C18 (4.6X 250mm, 5 μm);
mobile phase: methanol-phosphate buffer (taking 1.36g of monopotassium phosphate, adding water to dissolve and dilute the monopotassium phosphate into 1000ml, adding 4ml of triethylamine, and adjusting the pH value to 3.5 by using phosphoric acid) (75: 25);
column temperature: 35 ℃;
flow rate: 1.0 ml/min;
detection wavelength: 253 nm;
sample introduction volume: 20 μ l.
The implementation steps are as follows:
(1) preparation of dissolution medium: SDS2.00g, sodium dihydrogen phosphate 6.00g (7.80 g sodium dihydrogen phosphate dihydrate) and sodium hydroxide 0.90g were weighed and diluted to 1000ml with water.
(2) Putting 11.13mg and 11.35mg of flunarizine hydrochloride as reference into a 100ml measuring flask, adding 10ml of ethanol to dissolve, adding a dissolved medium to dilute to a scale, and shaking up. Precisely measuring 5ml, placing into a 50ml measuring flask, adding dissolution medium, diluting to scale, and shaking to obtain reference solution.
(3) Taking one capsule, placing into a dissolution cup filled with 600ml dissolution medium, and collecting 1ml filtrate after 30min as test solution.
(4) Precisely measuring 20 μ l of each of the reference solution and the sample solution, injecting into a liquid chromatograph, and recording chromatogram. A blank solvent control test was also performed. The content was calculated as peak area by external standard method.
The chromatogram of the test sample is shown in FIG. 4.
The above description is only an embodiment of the present invention, but the scope of the present invention is not limited thereto, and the present invention also includes a granular formulation and a tablet formulation, which can be identified by the method of the present invention, and any person skilled in the art can easily think of the changes or substitutions within the technical scope of the present invention, and the present invention shall be covered by the scope of the claims.
Claims (9)
1. A detection method for dissolution determination of flunarizine hydrochloride capsules is characterized by comprising the following steps: the method comprises the following steps: the dissolution method is a paddle method, 100 r/min, and sodium dodecyl sulfate is added as a dissolution medium; detecting the eluate by high performance liquid chromatography, and using octadecylsilane chemically bonded silica as filler; an ultraviolet detector with the detection wavelength of 251-255 nm; the column temperature is 25-45 ℃; the flow rate is 0.9-1.1 ml/min; the mobile phase is methanol and phosphate buffer solution, and the phosphate buffer solution is sodium dihydrogen phosphate solution.
2. The method for detecting the dissolution test of the flunarizine hydrochloride capsule according to claim 1, which is characterized in that: the concentration of the added sodium dodecyl sulfate is 0.01 percent to 1.0 percent (w/V), and the pH value is 6.8.
3. The method for detecting the dissolution test of the flunarizine hydrochloride capsule according to claim 1, which is characterized in that: sodium lauryl sulfate was added at a concentration of 0.4% (w/V).
4. The method for detecting the dissolution test of the flunarizine hydrochloride capsule according to claim 1, which is characterized in that: the ratio of mobile phase methanol to phosphate buffer was 75: 25.
5. The method for detecting the dissolution test of the flunarizine hydrochloride capsule according to claim 1, which is characterized in that: the sodium dihydrogen phosphate solution was 0.01mol/L sodium dihydrogen phosphate solution.
6. The method for detecting the dissolution test of the flunarizine hydrochloride capsule according to claim 1, which is characterized in that: to the phosphate buffer, 4ml of triethylamine was added per 1000ml of mobile phase.
7. The method for detecting the dissolution test of the flunarizine hydrochloride capsule according to claim 1, which is characterized in that: the phosphate buffer was adjusted to pH 3.5. + -. 0.05 with phosphoric acid.
8. The method for detecting the dissolution test of the flunarizine hydrochloride capsule according to claim 1, which is characterized in that: the detection wavelength was 253 nm.
9. The method for detecting the dissolution test of the flunarizine hydrochloride capsule according to claim 1, which is characterized in that: the column temperature was 35 ℃ and the flow rate was 1.0 ml/min.
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