CN109596728B - Method for measuring dissolution rate of acarbose tablets - Google Patents
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- CN109596728B CN109596728B CN201811471899.XA CN201811471899A CN109596728B CN 109596728 B CN109596728 B CN 109596728B CN 201811471899 A CN201811471899 A CN 201811471899A CN 109596728 B CN109596728 B CN 109596728B
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Abstract
The invention discloses a method for measuring dissolution rate of acarbose tablets, which solves the problems of low separation degree by adopting a C18 column, high loss by adopting an amino column chromatographic column, serious acarbose peak tailing, long analysis time, low sensitivity and high consumption of organic reagents in the prior art. The method for measuring the dissolution rate of the acarbose tablets adopts the high performance liquid chromatography to measure the dissolution rate test solution, and the chromatographic conditions are as follows: adopting a C18 column as a chromatographic column and phosphate buffer as a mobile phase A; acetonitrile is a mobile phase B; methanol is used as a mobile phase C for gradient elution. The method is simple, is simple and convenient to operate, can simultaneously meet the separation degree requirement of the test solution of four dissolution media, and has the advantages of shorter detection time, higher sensitivity and less consumption of organic reagents in the mobile phase.
Description
Technical Field
The invention belongs to the technical field of pharmaceutical analysis, and particularly relates to a method for determining dissolution rate of acarbose tablets.
Background
Acarbose (Acarbose), an α -glucosidase inhibitor for the treatment of type 2 diabetes, is a complex oligosaccharide consisting of four monosaccharides, the structure of which contains an unsaturated cyclitol. The alpha-glucosidase inhibitor binds to alpha-glucosidase by competing with oligosaccharides at the upper cell brush rim of the small intestine, thereby delaying hydrolytic absorption of carbohydrates and lowering blood glucose. Acarbose has the following structural formula:
acarbose tablets are the first clinical alpha-glucosidase inhibitor developed in the middle of 70 th century by Bayer company in Germany, and are sold under the trade name of Babesipine, and are also produced by a plurality of pharmaceutical factories at home at present. The research on the pharmacological action of the acarbose shows that the acarbose is particularly suitable for Chinese patients taking carbohydrate as a staple food, particularly middle-aged and elderly diabetic patients. At present, the on-market of acarbose in China needs to be evaluated by the consistency of the imitation drugs. The dissolution curve is a key index for measuring whether the quality of the solid preparation is consistent.
In the prior art, the acarbose analysis method mainly adopts a C18 column and an amino column. However, the separation effect of the C18 column is not good, and the separation degree requirement of the sample solution of the four dissolution media cannot be satisfied at the same time. So 2015 version of Chinese pharmacopoeia adopts amino column. And the existing acarbose analysis method of pharmacopoeia of various countries has multiple use of amino columns. However, the amino column is adopted for analysis, so that the chromatographic column has large loss, serious acarbose peak tailing, long analysis time, low sensitivity and large consumption of organic reagents.
Therefore, it is an urgent need to solve the problems of the art to provide a method for measuring dissolution rate of acarbose tablets, which can simultaneously satisfy the separation degree requirements of test solutions of four dissolution media, and has the advantages of shorter detection time, higher sensitivity and less organic reagent consumption in a mobile phase.
Disclosure of Invention
The technical problem solved by the invention is as follows: the provided method for measuring the dissolution rate of the acarbose tablets solves the problems of low separation degree by adopting a C18 column, large loss by adopting an amino column chromatographic column, serious acarbose peak tailing, long analysis time, low sensitivity and large consumption of organic reagents in the prior art.
The technical scheme adopted by the invention is as follows:
the invention relates to a method for measuring dissolution rate of acarbose tablets, which adopts high performance liquid chromatography to measure a dissolution rate test sample solution, wherein the chromatographic conditions are as follows: adopting a C18 column as a chromatographic column and phosphate buffer as a mobile phase A; acetonitrile is a mobile phase B; methanol is used as a mobile phase C, gradient elution is carried out according to the specification of the following table,
| time (min) | Mobile phase A (%) | Mobile phase B (%) | Mobile phase C (%) |
| 0.00 | 100 | 0 | 0 |
| 8.00 | 60-80 | 15-30 | 1-10 |
| 8.01 | 100 | 0 | 0 |
| 14.00 | 100 | 0 | 0 |
。
Preferably, the gradient elution is carried out as specified in the table below,
| time (min) | Mobile phase A (%) | Mobile phase B (%) | Mobile phase C (%) |
| 0.00 | 100 | 0 | 0 |
| 8.00 | 70 | 25 | 5 |
| 8.01 | 100 | 0 | 0 |
| 14.00 | 100 | 0 | 0 |
。
Further, the preparation method of the buffer salt solution comprises the following steps: 300mg of anhydrous sodium dihydrogen phosphate and 350mg of anhydrous disodium hydrogen phosphate are taken, dissolved in water and diluted to 1000 ml.
Further, the preparation method of the dissolution test solution comprises the following steps: taking acarbose tablets, adopting a slurry method, taking water, a hydrochloric acid solution with the pH value of 1.0, an acetate buffer solution with the pH value of 4.5 and a phosphate buffer solution with the pH value of 6.8 as dissolution media respectively, and taking dissolution liquid as a test solution at different time points respectively at the rotation speed of 50 r/min.
Further, the method adopts an ultraviolet detector, the detection wavelength is 210nm, the temperature of a chromatographic column is 10-12 ℃, and the flow velocity of a mobile phase is 1.8-2.2 ml/min.
Further, the chromatographic column is a water-resistant C18 column.
Further, the type of the water-resistant C18 column is a water-resistant AQ-C18 column.
Further, the method comprises the following steps:
step 1: preparation of a test solution: taking acarbose tablets, adopting a slurry method, taking water, a hydrochloric acid solution with the pH value of 1.0, an acetate buffer solution with the pH value of 4.5 and a phosphate buffer solution with the pH value of 6.8 as dissolution media respectively, and taking dissolution liquid as a test solution at different time points respectively at the rotation speed of 50 r/min;
step 2: preparing a reference substance solution: an appropriate amount of acarbose reference substance is precisely weighed, and dissolved and diluted to prepare a solution containing 50 mu g of acarbose reference substance in 1ml as a reference substance solution by respectively using water, a hydrochloric acid solution with a pH value of 1.0, an acetate buffer solution with a pH value of 4.5 and a phosphate buffer solution with a pH value of 6.8 as solvents;
and step 3: and (3) respectively injecting 50 mu l of each of the test solution and the reference solution into a liquid chromatograph, measuring according to the chromatographic conditions, and recording a chromatogram.
Compared with the prior art, the method has the following beneficial effects:
the method is simple, is simple and convenient to operate, can simultaneously meet the separation degree requirement of the test solution of four dissolution media, and has the advantages of shorter detection time, higher sensitivity and less consumption of organic reagents in the mobile phase.
Drawings
FIG. 1 is a high performance liquid chromatogram of an aqueous solution of acarbose from example 1;
FIG. 2 is a high performance liquid chromatogram of the acarbose hydrochloric acid solution of example 1 at pH 1.0;
FIG. 3 is a high performance liquid chromatogram of the acetate solution of acarbose pH4.5 of example 1;
FIG. 4 is a high performance liquid chromatogram of the phosphate solution of acarbose pH6.8 of example 1;
FIG. 5 is a high performance liquid chromatogram of the hydrochloric acid solution of acarbose pH1.0 at 20 ℃ in example 2 at a column temperature;
FIG. 6 is a high performance liquid chromatogram of an aqueous solution of acarbose from example 3;
FIG. 7 is a high performance liquid chromatogram of the hydrochloric acid solution of pH1.0 of example 4;
FIG. 8 is a high performance liquid chromatogram of the acarbose hydrochloric acid solution of example 4 at pH 1.0;
FIG. 9 is a diagram of HPLC analysis using an amino column performed according to the dissolution method of acarbose tablet quality standard in the 'Chinese pharmacopoeia' 2015 edition in example 5.
Detailed Description
The technical solution of the present invention will be further described with reference to the following specific examples, but the present invention is not limited to these examples.
Example 1
The embodiment provides a method for determining the dissolution rate of acarbose tablets, which specifically comprises the following steps:
preparation of dissolution medium:
preparing a hydrochloric acid solution dissolving medium with pH1.0: weighing 9ml of hydrochloric acid solution, adding purified water to dilute the hydrochloric acid solution to 1000ml, and shaking up the hydrochloric acid solution to obtain the compound.
② preparing an acetate buffer solution dissolution medium with pH4.5: 114ml of 2mol/L acetic acid solution is weighed, 2.99g sodium acetate is weighed, water is added for dissolution and dilution to 1000ml, and the mixture is shaken up to obtain the sodium acetate.
Preparation of phosphate buffer solution dissolution medium with pH6.8: weighing 6.8g of potassium dihydrogen phosphate, adding 112.0ml of 0.2mol/L sodium hydroxide solution, diluting with water, dissolving, diluting to 1000ml, and stirring.
Fourthly, ultrapure water (meeting the specification of the 2015 version of Chinese pharmacopoeia).
Preparation of a test solution: taking acarbose tablet (specification: 50mg), according to a dissolution determination method (second method of 0931 in the four headings of the Chinese pharmacopoeia 2015 edition), taking 900ml of hydrochloric acid solution with pH of 1.0 as a dissolution medium, rotating at 50 revolutions per minute, operating according to the method, taking 10ml of dissolution liquid after 30 minutes, filtering, and taking a subsequent filtrate as a test solution.
The same procedure was used to prepare a sample solution using pH4.5 acetate buffer, pH6.8 phosphate buffer, and ultrapure water as dissolution media.
Preparing a reference substance solution: precisely weighing appropriate amount of acarbose reference substance, dissolving in water, and diluting to obtain solution containing 50 μ g of acarbose per 1ml as reference substance solution; the reference solution is prepared in 2 parts by the same method.
The same procedure was followed to obtain a control solution using pH4.5 acetate buffer, pH6.8 phosphate buffer, and ultrapure water as dissolution media.
And (3) detection:
a detection instrument: shimadzu LC-2030C high performance liquid chromatograph
Chromatographic conditions are as follows: a chromatographic column: inertsustatin AQ-C18, 4.6mm × 250mm, 5 μm; mobile phase: phosphate buffer solution (anhydrous sodium dihydrogen phosphate 300mg and anhydrous disodium hydrogen phosphate 354mg, dissolved in water and diluted to 1000ml) is used as mobile phase A; acetonitrile is a mobile phase B; methanol as mobile phase C, elution as specified in table 1:
TABLE 1
| Time (min) | Mobile phase A (%) | Mobile phase B (%) | Mobile phase C (%) |
| 0.00 | 100 | 0 | 0 |
| 8.00 | 70 | 25 | 5 |
| 8.01 | 100 | 0 | 0 |
| 14.00 | 100 | 0 | 0 |
The column temperature is 10-12 ℃; the flow rate was 2.0ml per minute; the detection wavelength was 210 nm.
Precisely measuring 50 μ l of each of the reference solution and the sample solution, injecting into a chromatograph, and recording chromatogram. The chromatograms are shown in FIGS. 1-4, and the chromatographic data are shown in Table 2:
TABLE 2 HPLC analysis data table of acarbose dissolution liquid in four dissolution media
| Sample name | Retention time (min) | Number of theoretical plates | Tailing factor |
| Aqueous acarbose solutions | 5.623 | 19222 | 1.11 |
| Acarbose pH1.0 hydrochloric acid solution | 5.578 | 30857 | 1.16 |
| Acarbose acetate solution pH4.5 | 5.490 | 22478 | 1.11 |
| Acarbose phosphate solution pH6.8 | 5.493 | 19643 | 1.12 |
As can be seen from the above table, the method of the present invention can simultaneously satisfy the separation degree requirements of the test solution of the four dissolution media, and has the advantages of short detection time and good peak shape.
Example 2
This example provides a method for determining dissolution of acarbose tablets using the present invention.
The test solution of example 1, in which a hydrochloric acid solution of ph1.0 was used as the dissolution medium, was used as the test solution, and the control solution, in which a hydrochloric acid solution of ph1.0 was used as the dissolution medium, was used as the control solution. The measurement conditions were the same as in example 1 except that the column temperature was 20 ℃. The chromatogram obtained is shown in FIG. 5. The results show that the acarbose retention in this example is slightly shifted forward, the degree of separation is good and the peak shape is good.
Example 3
Preparation of a test solution: taking acarbose tablet (specification: 100mg), according to dissolution determination method (second method of 0931 in the four-part general rule of Chinese pharmacopoeia 2015 edition), taking 900ml of water as dissolution medium, rotating speed 50r/min, operating according to the method, taking 10ml of dissolution liquid after 30 min, filtering, and taking the subsequent filtrate as test solution.
Preparing a reference substance solution: precisely weighing appropriate amount of acarbose reference substance, dissolving in water, and diluting to obtain solution containing 50 μ g of acarbose per 1ml as reference substance solution; the reference solution is prepared in 2 parts by the same method.
And (3) determination:
chromatographic conditions are as follows: a chromatographic column: inertsustatin AQ-C18, 4.6mm × 250mm, 5 μm; mobile phase: phosphate buffer solution (anhydrous sodium dihydrogen phosphate 300mg and anhydrous disodium hydrogen phosphate 354mg, dissolved in water and diluted to 1000ml) is used as mobile phase A; acetonitrile-methanol (5:1) as mobile phase B, elution as specified in table 3:
TABLE 3
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0.00 | 100 | 0 |
| 8.00 | 70 | 30 |
| 8.01 | 100 | 0 |
| 14.00 | 100 | 0 |
The column temperature is 10-12 ℃; the flow rate was 2.0ml per minute; the detection wavelength is 210 nm; the injection volume was 50. mu.l.
Precisely measuring the sample solution, injecting 50 μ l each into chromatograph, and recording chromatogram. The chromatogram obtained is shown in FIG. 6. The results show that the gradient elution with the separation and proportional mixing of the methanol acetonitrile in the organic phase in the analytical method is equivalent.
Example 4
This example is a comparative example.
Preparation of dissolution medium:
preparation of hydrochloric acid solution dissolution medium with pH 1.0: weighing 9ml of hydrochloric acid solution, adding purified water to dilute the hydrochloric acid solution to 1000ml, and shaking up the hydrochloric acid solution to obtain the compound.
Preparation of a test solution: taking acarbose tablet (specification: 100mg), according to a dissolution determination method (second method of 0931 in the four-part general rule of Chinese pharmacopoeia 2015 edition), taking 900ml of hydrochloric acid solution with pH of 1.0 as a dissolution medium, rotating at 50 revolutions per minute, operating according to the method, taking 10ml of dissolution liquid after 30 minutes, filtering, and taking a subsequent filtrate as a test solution.
Chromatographic conditions (reference: determination of dissolution of acarbose tablet: Wufang, von Konmin, Caotong, Wanglianzi, Zihuijian, Li Huimin, northwest pharmaceutical journal, 2017, stage 01)
A chromatographic column: YMC-Pack ODS-AQ, 4.6 mm. times.150 mm, 5 μm
Mobile phase: dissolving in phosphate buffer (anhydrous sodium dihydrogen phosphate 279mg and potassium dihydrogen phosphate 600mg in water 100ml, and diluting to 1000ml) -acetonitrile (93:7)
The column temperature was 40 ℃; flow rate was 1.5ml per minute; the detection wavelength is 210 nm; the injection volume was 50. mu.l.
Precisely measuring the sample solution and 50 μ l of hydrochloric acid solution with pH of 1.0, injecting into chromatograph, and recording chromatogram, wherein the obtained chromatogram is shown in figures 7-8.
The results show that the acarbose peak in the test solution is too early, the retention time is less than 2 minutes, and the dissolution medium ph1.0 hydrochloric acid solution interferes the detection of the acarbose peak in the test solution. This method is not suitable for dissolution determination of acarbose tablets.
Example 5
This example is a comparative example.
This example was carried out according to the acarbose dissolution method recorded in the Chinese pharmacopoeia (2015 second division), and the chromatogram obtained is shown in FIG. 9. As can be seen from FIG. 9, the acarbose peak was retained for a long time, and the peak of the chromatogram was severely smeared.
The above-mentioned embodiment is only one of the preferred embodiments of the present invention, and should not be used to limit the scope of the present invention, but all the insubstantial modifications or changes made within the spirit and scope of the main design of the present invention, which still solve the technical problems consistent with the present invention, should be included in the scope of the present invention.
Claims (2)
1. A method for measuring dissolution rate of acarbose tablets is characterized in that a dissolution rate test sample solution is measured by adopting a high performance liquid chromatography, and the chromatographic conditions are as follows: a water-resistant AQ-C18 column is adopted as a chromatographic column, and a phosphate buffer solution is adopted as a mobile phase A; acetonitrile is a mobile phase B; methanol is used as a mobile phase C, gradient elution is carried out according to the specification of the following table,
The method adopts an ultraviolet detector, the detection wavelength is 210nm, the temperature of a chromatographic column is 10-12 ℃, and the flow rate of a mobile phase is 1.8-2.2 ml/min;
the preparation method of the phosphate buffer solution comprises the following steps: dissolving 300mg of anhydrous sodium dihydrogen phosphate and 350mg of anhydrous disodium hydrogen phosphate in water, and diluting to 1000 ml;
the preparation method of the dissolution test solution comprises the following steps: taking acarbose tablets, adopting a slurry method, taking water, a hydrochloric acid solution with the pH value of 1.0, an acetate buffer solution with the pH value of 4.5 and a phosphate buffer solution with the pH value of 6.8 as dissolution media respectively, and taking dissolution liquid as a test solution at different time points respectively at the rotation speed of 50 r/min.
2. The method for determining the dissolution rate of acarbose tablets according to claim 1, which comprises the following steps:
step 1: preparation of a test solution: taking acarbose tablets, adopting a slurry method, taking water, a hydrochloric acid solution with the pH value of 1.0, an acetate buffer solution with the pH value of 4.5 and a phosphate buffer solution with the pH value of 6.8 as dissolution media respectively, and taking dissolution liquid as a test solution at different time points respectively at the rotation speed of 50 r/min;
step 2: preparing a reference substance solution: an appropriate amount of acarbose reference substance is precisely weighed, and dissolved and diluted to prepare a solution containing 50 mu g of acarbose reference substance in 1ml as a reference substance solution by respectively using water, a hydrochloric acid solution with a pH value of 1.0, an acetate buffer solution with a pH value of 4.5 and a phosphate buffer solution with a pH value of 6.8 as solvents;
and step 3: and (3) respectively injecting 50 mu l of each of the test solution and the reference solution into a liquid chromatograph, measuring according to the chromatographic conditions, and recording a chromatogram.
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| CN113325101B (en) * | 2021-05-27 | 2022-09-16 | 南京海纳医药科技股份有限公司 | Method for detecting dissolution rates of glucosamine hydrochloride preparation in different media |
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| STABILITY INDICATING RPHPLC METHOD DEVELOPMENT AND VALIDATION OF SIMULTANEOUS ESTIMATION OF METFORMIN AND ACARBOSE IN BULK AND PHARMACEUTICAL FORMULATION;Alagar Raja.M et al.;《Asian Journal of Research in Biological and Pharmaceutical Sciences》;20151231;第3卷(第2期);全文 * |
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Denomination of invention: A method for determining the dissolution of Acarbose Tablets Effective date of registration: 20220531 Granted publication date: 20220208 Pledgee: Bank of Chengdu science and technology branch of Limited by Share Ltd. Pledgor: CHENGDU SINO-STRONG PHARMACEUTICAL Co.,Ltd. Registration number: Y2022980006615 |
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Date of cancellation: 20221228 Granted publication date: 20220208 Pledgee: Bank of Chengdu science and technology branch of Limited by Share Ltd. Pledgor: CHENGDU SINO-STRONG PHARMACEUTICAL Co.,Ltd. Registration number: Y2022980006615 |