Determination method for sampling detection of metformin hydrochloride oral solution content in hospital

Abstract

The invention belongs to the field of medicine and is used for measuring the medicine content of a metformin hydrochloride oral solution in hospitals. The invention provides a method for detecting the content of a metformin hydrochloride oral solution, which is a high performance liquid chromatography (HPLC), and the HPLC conditions are as follows: the chromatographic column is an Agilent C18 chromatographic column, the column temperature is 20-40 DEG C, the mobile phase is that 0.5-1.5 g of lauryl sodium sulfate is dissolved in 620 ml of dilute phosphoric acid, 200-400 ml of acetonitrile, the flow rate is 0.5-1.5 mL / min, the wavelength is 230 nm, and the sample injection volume is 5-20 [mu] L. According to the method disclosed by the invention, the interference of auxiliary materials can be eliminated, the main content of the medicine can be rapidly determined, the content detection peak appearance time is before 7 minutes (min) (the peak appearance time is 6.232 minutes), and the detection efficiency is remarkably improved; in addition, the detection method has the advantages of simplicity in operation, high sensitivity, wide linear range, good accuracy and the like.

Description

Technical field:

[0001] The invention belongs to the field of medicine, and in particular relates to a quantitative detection method for drug sampling and content determination of metformin hydrochloride oral solution in hospitals. Background technology:

[0002] Metformin hydrochloride is one of the classic hypoglycemic drugs. It is an oral hyperglycemic drug for the treatment of type 2 diabetes, especially the first choice for obese and hyperinsulinemia. Its main pharmacological action is to promote the anaerobic degradation of sugar during sugar metabolism and increase the uptake of glucose by peripheral tissues such as muscle and fat. At present, the mainstream dosage forms in China are solid preparations such as tablets, capsules, and sustained-release tablets. Metformin hydrochloride oral solution preparations are mainly used in some children's hospitals in China. It has obvious advantages in clinical practice and can well solve the problems of poor compliance and difficult dosage of special populations such as children. However, due to the short time on the market in China and the low market penetration rate, there is no mature methodologically verified content determination method for the determination of this oral solution dosage form. The method of the present invention can effectively solve the urgent need for sampling content detection of metformin hydrochloride oral solution products in hospitals at present. Summary of the invention:

[0003] The present invention aims to provide a method for determining the content of metformin hydrochloride oral solution with simple operation, high detection efficiency and high accuracy, which is used for sample sampling and detection in hospitals. The peak time of main component content detection is 7 minutes (min) ago (the peak time is 6.232 min), that is, a batch of drugs can be determined within 7 minutes after the sample is prepared, meeting the hospital's demand for high-efficiency sampling and detection.

[0004] The following invention is thus provided:

[0005] The present invention belongs to the field of medicine and is used for determining the drug content of metformin hydrochloride oral solution in hospitals. The present invention provides a method for detecting the content of metformin hydrochloride oral solution, which is an HPLC method, and the HPLC conditions are as follows:

[0006] Chromatographic column: Agilent C18 column;

[0007] Wavelength: 230nm;

[0008] Column temperature: 20℃-40℃;

[0009] Flow rate 0.5mL / min-1.5mL / min;

[0010] Injection volume: 5-20 μL;

[0011] Mobile phase: 0.5g-1.5g sodium dodecyl sulfate dissolved in 620mL dilute phosphoric acid: 200mL-400mL acetonitrile; wherein, the HPLC method for determining the content of the main component metformin hydrochloride has been verified by methodology, and the interference of auxiliary materials can be eliminated.

[0012] In one or more embodiments of the present invention, the chromatographic column is an Agilent C18 chromatographic column. Preferably, the chromatographic column parameters are 250 mm x 4.6 mm, 5.0 μm or 150 mm x 4.6 mm, 5.0 μm or 250 mm x 4.6 mm, 10.0 μm.

[0013] In one or more embodiments of the present invention, the detection method, wherein the column temperature is 20°C-40°C, 30°C-40°C or 30°C.

[0014] In one or more embodiments of the present invention, the detection method, wherein the flow rate is 0.5mL / min-1.5ml / min, 1.0mL / min-1.5mL / min or 1.5mL / min.

[0015] In one or more embodiments of the present invention, the detection method, wherein the fluidity is 0.5g-1.5g sodium dodecyl sulfate dissolved in 620mL dilute phosphoric acid: 200mL-400mL acetonitrile, 1.0g-1.5g sodium dodecyl sulfate dissolved in 620mL dilute phosphoric acid: 200mL-400mL acetonitrile, 1.5g sodium dodecyl sulfate dissolved in 620mL dilute phosphoric acid: 200mL-400mL acetonitrile or 0.5g-1.5g sodium dodecyl sulfate dissolved in 620mL dilute phosphoric acid: 400mL acetonitrile.

[0016] In one or more embodiments of the present invention, the detection method, wherein the injection volume is: 5-20 μL, 10-20 μL or 20 μL. Description of the drawings:

[0017] Figure 1 : Sampling and testing the peak time of the main component of metformin hydrochloride oral solution samples;

[0018] Figure 2 : Sampling test of metformin hydrochloride oral solution sample excipient (saccharin calcium) peak time chart;

[0019] Figure 3 : Sampling test of metformin hydrochloride oral solution sample excipient (xylitol) peak time diagram;

[0020] Figure 4 : Sampling test of metformin hydrochloride oral solution sample excipients (solvent) peak time diagram;

[0021] Figure 5 : Sampling test of metformin hydrochloride oral solution samples and determination of peak time of related substances. Specific implementation method:

[0022] The present invention is described in detail below in conjunction with examples, but the following examples are only preferred implementations of the present invention, and the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field within the technical scope disclosed in the present invention, according to the technical solution and inventive concept of the present invention, make equivalent replacements or changes, which should be covered within the protection scope of the present invention.

[0023] In the following examples, the main instruments, reagents and materials used are as follows: high performance liquid chromatograph (Agilent 1100), metformin hydrochloride reference substance (China Food and Drug Inspection Institute), metformin hydrochloride oral solution (batch number: H180257A, manufacturer: Mikart, Inc), sodium dodecyl sulfate (Hebei Bailingwei Ultrafine Materials Co., Ltd.), acetonitrile (Fisher, USA), phosphoric acid (Sinopharm Chemical Reagent Co., Ltd.).

[0024] Sample preparation:

[0025] ① Reference solution: 20 mg of metformin hydrochloride reference substance is accurately weighed and quantitatively diluted with 40% acetonitrile aqueous solution to prepare a solution containing approximately 100 μg of metformin hydrochloride per 1 mL, which is used as the reference solution.

[0026] ② Test solution: Accurately measure 5 mL of this product (equivalent to 500 mg of metformin hydrochloride), add 40% acetonitrile aqueous solution to quantitatively dilute it to make a solution containing approximately 100 μg of metformin hydrochloride per 1 mL as the test solution.

[0027] Implementation Case 1:

[0028] 1. Test methods

[0029] Chromatographic column: Agilent C18 column (4.6 mm × 150 mm; 5 μm);

[0030] Wavelength: 230nm;

[0031] Column temperature: 40°C;

[0032] Flow rate: 1.5 mL / min;

[0033] Injection volume: 20 μl;

[0034] Mobile phase: sodium dodecyl sulfate solution (1.0 g sodium dodecyl sulfate dissolved in 620 mL dilute phosphoric acid (1 mL-2500 mL): acetonitrile = 62:38

[0035] 2. Detection and sampling

[0036] Accurately measure 20 μL of the reference solution and the test solution, inject them into a high performance liquid chromatograph (HPLC), record the chromatogram, and calculate the content by peak area using the external standard method.

[0037] 3. Content calculation

[0038] The content was calculated by peak area using the external standard method. The calculation formula is:

[0039]

[0040]

[0041] Note: A1 is the average peak area of ​​five consecutive injections of reference solution 1

[0042] A2 is the average peak area of ​​two consecutive injections of reference solution 2

[0043] A is the average peak area of ​​the reference solution after 2 conversion

[0044] A is the main peak area in the test solution

[0045] C1 is the concentration of reference solution 1, μg / mL

[0046] C2 is the concentration of reference solution 2, μg / mL

[0047] Labelled amount: 100mg / mL

[0048] 4. Measurement results:

[0049]

[0050] Implementation Case 2:

[0051] 1. Test methods

[0052] Chromatographic column: Agilent C18 column (4.6 mm × 150 mm; 5 μm);

[0053] Wavelength: 230nm;

[0054] Column temperature: 30°C;

[0055] Flow rate: 1.0 ml / min;

[0056] Injection volume: 5 μL;

[0057] Mobile phase: sodium dodecyl sulfate solution (1.0 g sodium dodecyl sulfate dissolved in 620 mL dilute phosphoric acid (1 mL-2500 mL): acetonitrile = 62:38

[0058] 2. Detection and sampling

[0059] Accurately measure 10 μL of the reference solution and the test solution, inject them into a high performance liquid chromatograph (HPLC), record the chromatogram, and calculate the content by the peak area using the external standard method.

[0060] 3. Content calculation

[0061] The content was calculated by peak area using the external standard method. The calculation formula is:

[0062]

[0063]

[0064] Note: A1 is the average peak area of ​​five consecutive injections of reference solution 1

[0065] A2 is the average peak area of ​​two consecutive injections of reference solution 2

[0066] A is the average peak area of ​​the reference solution after 2 conversion

[0067] A is the main peak area in the test solution

[0068] C1 is the concentration of reference solution 1, μg / mL

[0069] C2 is the concentration of reference solution 2, μg / mL

[0070] Labelled amount: 100mg / mL

[0071] 4. Measurement results

[0072]

[0073] Implementation Case 3

[0074] 1. Test methods

[0075] Chromatographic column: Agilent C18 column (4.6 mm × 150 mm; 5 μm);

[0076] Wavelength: 230nm;

[0077] Column temperature: 20°C;

[0078] Flow rate: 1.5 ml / min;

[0079] Injection volume: 5 μL;

[0080] Mobile phase: sodium dodecyl sulfate solution (1.0 g sodium dodecyl sulfate dissolved in 620 mL dilute phosphoric acid (1 mL-2500 mL): acetonitrile = 62:38

[0081] 2. Detection and sampling

[0082] Accurately measure 15 μL of the reference solution and the test solution, inject them into a high performance liquid chromatograph (HPLC), record the chromatogram, and calculate the content by peak area using the external standard method.

[0083] 3. Content calculation

[0084] The content was calculated by peak area using the external standard method. The calculation formula is:

[0085]

[0086]

[0087] Note: A1 is the average peak area of ​​five consecutive injections of reference solution 1

[0088] A2 is the average peak area of ​​two consecutive injections of reference solution 2

[0089] A is the average peak area of ​​the reference solution after 2 conversion

[0090] A is the main peak area in the test solution

[0091] C1 is the concentration of reference solution 1, μg / mL

[0092] C2 is the concentration of reference solution 2, μg / mL

[0093] Labelled amount: 100mg / mL

[0094] 4. Measurement results

[0095]

[0096] Implementation case 4: Method validation of content determination

[0097] 1. Solution preparation

[0098] ① Preparation of linear relationship investigation solution: take about 20 mg of metformin hydrochloride reference substance, accurately weigh it, put it in a 100 mL volumetric flask, add 40% acetonitrile aqueous solution to dissolve and dilute to the scale, shake well, and use it as a stock solution; take the stock solution and add 40% acetonitrile aqueous solution to dilute to the concentration of 160 μg / mL, 120 μg / mL, 100 μg / mL, 80 μg / mL, and 50 μg / mL as linear solutions; take the linear solution and measure it according to the law, and perform linear regression with the sample concentration as the horizontal axis and the peak area as the vertical axis. The test results are shown in Tables 1 and Figure 1 as shown.

[0099] ②Durability test solution preparation: Investigate the influence of chromatographic conditions (flow rate and column temperature) fluctuation on the content determination results. The parameters of chromatographic condition changes are shown in the table below. Under the conditions of flow rate and column temperature changes, the RSD of the content change of the test solution is less than 2.0%. The durability results are shown in Table 2.

[0100] ③ Preparation of system adaptability solution: Weigh an appropriate amount of metformin hydrochloride sample, place it in a 100mL volumetric flask, dissolve and dilute it with a diluent to make a 0.2mg / mL stock solution; accurately measure 5mL of the stock solution and place it in a 10mL volumetric flask, add a diluent to the scale, shake well, filter with a 0.22μm microporous membrane, accurately measure and determine according to the law, the system adaptability results are shown in Table 3.

[0101] ④ Accuracy test solution preparation: accurately weigh 20 mg of metformin hydrochloride reference substance, dilute it with 40% acetonitrile aqueous solution (diluent) to 1 mg / mL solution as stock solution; transfer 0.8 mL, 1.0 mL, and 1.2 mL of stock solution to 10 mL volumetric flask respectively, dilute to scale with diluent, and shake well; prepare 3 samples of different concentrations in the same way, and filter with 0.22 μm microporous filter membrane. Accurately measure and determine according to the law, calculate the recovery rate, and the accuracy test results are shown in Table 4.

[0102] ⑤ Preparation of solution for precision investigation: Accurately measure an appropriate amount of metformin hydrochloride oral solution and dilute it with 40% acetonitrile aqueous solution to a concentration of 100 μg / mL as the test solution. Prepare 6 test solutions in the same way and determine them according to the method. The results of the precision investigation are shown in Table 5.

[0103] 2. Test methods

[0104] Chromatographic column: Agilent C18 column (4.6 mm × 150 mm; 5 μm);

[0105] Wavelength: 230nm;

[0106] Column temperature: 40°C;

[0107] Flow rate: 1.5 ml / min;

[0108] Injection volume: 20 μL;

[0109] Mobile phase: sodium dodecyl sulfate solution (1.0 g sodium dodecyl sulfate dissolved in 620 mL dilute phosphoric acid (1 mL-2500 mL): acetonitrile = 62:38

[0110] 3. Test results

[0111] Table 1 Results of linear relationship test of detection method

[0112]

[0113] Table 2 Durability test results of the detection method

[0114]

[0115]

[0116] Table 3 Results of system suitability test of detection method

[0117]

[0118] Table 4 Results of the accuracy test of the detection method

[0119]

[0120] Table 5 Results of precision test of detection method

[0121]

[0122]

[0123] The above results show that the present invention is not only highly efficient in detection (peak time of the main component: 6.232 min), simple and quick to operate, but also has accurate measurement results, a wide linear range, and high sensitivity. It is a simple and feasible analytical method for sampling and determining the content of metformin hydrochloride oral solution in hospitals.

Claims

1. A method for sampling and detecting the content of metformin hydrochloride oral solution in a hospital, which is an HPLC method, wherein the HPLC conditions are as follows: Chromatographic column: Agilent C18 column; Wavelength: 230nm; Column temperature: 20℃-40℃; Flow rate 0.5mL / min-1.5mL / min; Injection volume: 5-20 μL; Mobile phase: 0.5g-1.5g sodium dodecyl sulfate dissolved in 620ml dilute phosphoric acid: 200mL-400mL acetonitrile; in, The HPLC method for determining the content of the main ingredient metformin hydrochloride has been validated by methodology and can eliminate the interference of auxiliary materials.

2. A method for determining the content of metformin hydrochloride oral solution for sampling in a hospital according to claim 1, characterized in that An Agilent C18 column was used with a wavelength of 230 nm and a column temperature of 20°C-40°C. The mobile phase was a mixture of 0.5 g-1.5 g sodium dodecyl sulfate dissolved in 620 ml of dilute phosphoric acid: 200 mL-400 mL of acetonitrile.

3. According to the method described in claim 1 or 2, this invention is characterized in that the sampling content determination of the metformin hydrochloride oral solution in the hospital can be carried out for quantitative analysis.

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