Method for detecting residual contents of olmesartan medoxomil, olmesartan and hydrochlorothiazide in granulator

Abstract

The invention discloses a method for detecting the content of residual olmesartan medoxomil, olmesartan and hydrochlorothiazide in a granulator, which is characterized in that an HPLC (High Performance Liquid Chromatography) method is used for detection, and an Agilent Zorbax Rx C8 chromatographic column with octyl silane bonded silica gel as a filler, 4.6 mm * 15 cm and 5 mu m is used; a solution with the volume ratio of acetonitrile to a phosphate buffer solution being 20: 80 is used as a mobile phase A, a solution with the volume ratio of acetonitrile to the phosphate buffer solution being 80: 20 is used as a mobile phase B, and linear gradient elution is carried out. The method is suitable for detection of residues at a relatively low concentration level, is short in time consumption and is suitable for daily monitoring scenes of production equipment.

Description

Technical Field

[0001] The invention relates to the field of drug analysis, and in particular to a method for detecting the contents of residual olmesartan medoxomil, olmesartan and hydrochlorothiazide in a granulator. Background Art

[0002] Different types of drugs may need to share production equipment during the production process. According to regulations, the cleaning effect of the equipment needs to be evaluated after the production is completed to ensure that no drug components remain in the equipment and that they are not mixed with other types of drugs to become impurities.

[0003] Generally speaking, the specific cleaning residue limit is calculated based on the different types of drugs produced on the same line. Changes in production line varieties and additions will cause changes in residue limits, which may cause detection methods with a small concentration range to exceed the quantifiable range. In many cases, the analytical methods used to evaluate the cleaning effect of equipment need to have higher sensitivity and lower detection limits than general analytical methods, which may cause the existing general analytical methods to fail to meet the requirements of cleaning effect evaluation.

[0004] The residues that may appear in the equipment currently used to produce Olmesartan Medoxomil and Hydrochlorothiazide Tablets are Olmesartan Medoxomil, Olmesartan and Hydrochlorothiazide. Olmesartan comes from the degradation of Olmesartan Medoxomil. In the prior art, there are reports of relevant analytical methods, but they all have some defects, such as the lower limit of the measured concentration is not low enough, the analysis time is too long, etc., which cannot meet the needs of equipment cleaning effect evaluation. The concentration of the compound is usually at a trace or even trace level; the detection method using the content determination type is not sensitive enough to accurately quantify the trace compounds, and the compounds that can be detected are limited. In an article published in the 11th issue of Volume 18 of the Chinese Pharmacist in 2015, Liu Yanming et al. disclosed a HPLC method for simultaneously determining the contents of three ingredients, namely, olmesartan medoxomil, amlodipine and hydrochlorothiazide, in compound olmesartan medoxomil tablets. However, the lower limit of the concentration that can be detected by this method is as follows: olmesartan medoxomil: 4 μg / ml to 80 μg / ml, hydrochlorothiazide: 1.25 μg / ml to 25 μg / ml, and amlodipine: 1 μg / ml to 20 μg / ml. The detection concentration is not low enough, and the running time is about 25 minutes, so there is still room for further improvement.

[0005] In an article published in "China Pharmacy" Vol. 31, No. 7, 2020, Li Haoxian and other authors disclosed a HPLC method for determining the content of four related substances in olmesartan medoxomil and hydrochlorothiazide preparations. The lower limit of the detectable concentration is: Olmesartan medoxomil: 0.2527~7.5800μg / ml Hydrochlorothiazide: 1.1521~4.5629μg / ml. The detection concentration is not low enough, and the types of substances detected are only olmesartan medoxomil and hydrochlorothiazide. The running time is as long as 80 minutes, which cannot meet the cleaning validation requirements for the production of equipment containing two ingredients: olmesartan medoxomil and hydrochlorothiazide. Summary of the invention

[0006] Based on the research background, this paper proposes an analytical method for verifying the cleaning effect of Olmesartan Medoxomil and Hydrochlorothiazide Tablets production equipment. Compared with the existing technology, this method can simultaneously detect the three components of Olmesartan, Olmesartan Medoxomil and Hydrochlorothiazide remaining in the instrument, with a detection limit concentration as low as 10 -2 μg / ml level, and one analysis takes only 7 minutes, which is suitable for the detection scenario of actual drug production, making up for the defects of insufficient detection concentration lower limit and long analysis time caused by direct conversion of existing technologies. The method of the present invention can detect compounds with extremely low concentrations, has a wide range of detection components and a wide concentration range (quantifiable range), which is very necessary for the detection of cleaning residues. The chromatographic conditions of this method are: using a C8 column and gradient elution, and the specific steps are: Sampling: Place a cotton swab in a 100ml beaker, add 50ml ethanol to clean it, take it out and shake it until there are no droplets, then use the cotton swab to wipe the sampling area of ​​the equipment for sampling. The sampling area size is 25 square centimeters. Then place the cotton swab head in a test tube, add 10.0ml of 50% acetonitrile solution, ultrasonically treat for 10 minutes, centrifuge, and take the supernatant to obtain the test solution; Solution preparation: Reference solution: Olmesartan medoxomil concentration is 7.5 μg / mL, hydrochlorothiazide concentration is 1.1 μg / mL in 50% acetonitrile solution; Olmesartan test solution: Olmesartan concentration is 7.0 μg / ml in 50% acetonitrile solution; Phosphate buffer: 1.36 g / L potassium dihydrogen phosphate solution, adjust the pH to 2.5 with phosphoric acid; Chromatographic injection: The chromatographic column was Agilent Zorbax Rx C8, 4.6 mm × 15 cm, 5 μm, with octylsilane bonded silica as the filler; the mobile phase A was a solution of acetonitrile: phosphate buffer in a volume ratio of 20:80, and the mobile phase B was a solution of acetonitrile: phosphate buffer in a volume ratio of 80:20, and linear gradient elution was performed; the flow rate was 1.5 ml per minute; the detection wavelength was 225 nm; the column temperature was 30 ° C; the injector temperature was 5 ° C; the injection volume was 10 μl, and the elution gradient was:

[0007] Solution preparation: Diluent: 50% acetonitrile Blank solution: diluent Wiping fluid: 95% ethanol Blank cotton swab solution: Place a cotton swab (moisten the swab with wiping solution) in a test tube, add 10.0 mL of diluent, ultrasonically treat for 10 min, centrifuge, and take the supernatant.

[0008] Blank excipient stock solution: Take about 325 mg of blank excipient, accurately weigh, place in a 100 mL volumetric flask, dissolve with diluent, ultrasonically treat for 10 min, dilute to the scale, shake well, and obtain.

[0009] Olmesartan medoxomil reference stock solution: Take about 15.0 mg of Olmesartan medoxomil reference solution, accurately weigh it, place it in a 100 mL volumetric flask, dissolve it in acetonitrile, dilute it to the mark, shake well, and you have it. (Concentration is about: 150.0 μg / mL) Hydrochlorothiazide reference substance stock solution: Take about 5.5 mg of hydrochlorothiazide reference substance, accurately weigh it, place it in a 100 mL volumetric flask, dissolve it in acetonitrile, dilute it to the mark, shake it well, and you have it. (Concentration is about: 55.0 μg / mL) Reference solution: Accurately pipette 5.0 mL of Olmesartan Medoxomil Reference Solution and 2.0 mL of Hydrochlorothiazide Reference Solution into a 100 mL volumetric flask, dilute to scale with diluent, shake well, and the solution is ready. Prepare two parallel samples in the same way. (The concentration of Olmesartan Medoxomil is about 7.5 μg / mL, and the concentration of Hydrochlorothiazide is about 1.1 μg / mL.) (Stable for 51 hours at room temperature and 5°C) Test solution stock solution: Take about 37.5 mg of Olmesartan Medoxomil reference substance, about 5.5 mg of Hydrochlorothiazide reference substance and about 325 mg of blank excipient, accurately weigh them, place them in a 100 mL volumetric flask, dissolve them with diluent, ultrasonicate for 10 minutes, dilute to the scale with diluent, shake well, and you have it. (The concentration of Olmesartan Medoxomil is about 375.0 μg / mL, and the concentration of Hydrochlorothiazide is about 55.0 μg / mL.) Olmesartan test solution stock solution: take about 7.0 mg of Olmesartan reference substance and about 65 mg of blank excipient, accurately weigh them, put them in a 20 ml volumetric flask, dissolve them with diluent, ultrasonicate for 10 min, dilute to the scale with diluent, shake well, and you have it. (The concentration of Olmesartan is about 350.0 μg / ml.) Test solution: Take a simulation device made of stainless steel plate, take a 25cm2 area, drop 0.2mL of test solution stock solution into the area, blow dry, wipe the area with two cotton swabs, cut off the two cotton swabs after wiping, place them in the same test tube, add 10.0mL of diluent, ultrasonically treat for 10 minutes, centrifuge, and take the supernatant. (The concentration of olmesartan medoxomil is about 7.5μg / mL, and the concentration of hydrochlorothiazide is about 1.1μg / mL.) (Stable for 14 hours at room temperature and 5℃) Olmesartan test solution: Take a simulation device made of stainless steel plate, take a 25cm2 area, drop 0.2ml of Olmesartan test solution stock solution in the area, blow dry, wipe the area with two cotton swabs, cut off the two cotton swabs after wiping, place them in the same test tube, add 10.0ml of diluent, ultrasonically treat for 10 minutes, centrifuge, and take the supernatant. (The concentration of Olmesartan is about 7.0μg / ml.) Blank wiping solution: Take a simulated device made of stainless steel plate, take an area of ​​25cm2, and wipe the above area with two cotton swabs (moistened with 95% ethanol and squeezed until there are no droplets when using). After wiping, cut off the heads of the two cotton swabs and place them in the same test tube, add 10.0mL of diluent, ultrasonically treat for 10 minutes, centrifuge, and take the supernatant.

[0010] Injection procedure: After the system is stable, inject 1-3 injections of blank solution, 1 injection of blank cotton swab solution, 1 injection of blank excipient solution, 1 injection of blank wipe solution, 5 injections of control solution 1, 1 injection of control solution 2, and 1 injection of test solution each. In the sequence, inject 1 reference solution for back calibration every 8 hours and at the end, and record the chromatogram.

[0011] calculate: Olmesartan result value (μg) correction factor method = R1 / Rs*F*Cs×10 Where: R U : Peak area of ​​olmesartan medoxomil in the test solution; R S : The peak area of ​​olmesartan medoxomil of the control solution; C S : The concentration of olmesartan medoxomil in the control solution (μg / ml); 10: The dilution factor of the test solution; R1: peak area of ​​Olmesartan in the test solution; F: correction factor of Olmesartan relative to Olmesartan Medoxomil; Olmesartan residue (mg) = Olmesartan result value (mg) / Olmesartan minimum sampling recovery rate Olmesartan Medoxomil result value (μg) = R U / R S ×C S ×10 Where: R U : peak area of ​​the test solution; R S : Peak area of ​​control solution; C S : Concentration of control solution (μg / ml); 10: dilution factor of test solution.

[0012] Olmesartan medoxomil residue (mg) = Olmesartan medoxomil result value (mg) / Olmesartan medoxomil minimum sampling recovery rate Comprehensive result (in terms of Olmesartan Medoxomil, mg) = Olmesartan residue * 558.59 / 446.5 + Olmesartan residue Among them: 558.59: molecular weight of Olmesartan Medoxomil; 446.5: molecular weight of Olmesartan.

[0013] Hydrochlorothiazide result value (μg) = R U / R S ×C S ×10 Where: R U : Measured value of the test solution; R S : The measured value of the control solution; C S : Concentration of hydrochlorothiazide control solution (μg / ml); 10: dilution factor of test solution.

[0014] Hydrochlorothiazide residue (mg) = hydrochlorothiazide result value (mg) / minimum sampling recovery rate of hydrochlorothiazide Recovery rate %= in: A 2 ——Peak area of ​​reference solution 2; M 1 ——The sampling volume of reference solution 1 (unit: mg); A 1 ——Average peak area of ​​reference solution 1; M 2 ——The sampling volume of reference solution 2 (unit: mg).

[0015] Method validation results:

[0016] Beneficial effect: Compared with the existing technology, this solution can simultaneously detect the three components of Olmesartan, Olmesartan Medoxomil and Hydrochlorothiazide remaining in the granulator, with a detection limit concentration as low as 10 -2 The concentration is at the μg / ml level and one analysis takes only 7 minutes, which is suitable for the actual drug production detection scenarios. It makes up for the shortcomings of the existing technology such as insufficient detection concentration limit and long analysis time. DETAILED DESCRIPTION Example

[0018] System suitability experiment Dilution / blank solution: 50% acetonitrile Olmesartan medoxomil reference stock solution: Take about 15.0 mg of Olmesartan medoxomil reference solution, accurately weigh it, place it in a 100 mL volumetric flask, dissolve it in acetonitrile, dilute it to the mark, shake well, and you have it. (Concentration is about: 150.0 μg / mL) Hydrochlorothiazide reference substance stock solution: Take about 5.5 mg of hydrochlorothiazide reference substance, accurately weigh it, place it in a 100 mL volumetric flask, dissolve it in acetonitrile, dilute it to the mark, shake it well, and you have it. (Concentration is about: 55.0 μg / mL) Reference solution: Accurately pipette 5.0 mL of Olmesartan Medoxomil Reference Solution and 2.0 mL of Hydrochlorothiazide Reference Solution into a 100 mL volumetric flask, dilute to scale with diluent, shake well, and the solution is ready. Prepare two parallel samples in the same way. (The concentration of Olmesartan Medoxomil is about 7.5 μg / mL, and the concentration of Hydrochlorothiazide is about 1.1 μg / mL.)

[0019]

[0020] After the system is stable, inject 1-3 injections of blank solution, 5 injections of control solution 1, 1 injection of reference solution 2, and return to calibration by injecting reference solution 1 every 8 hours and at the end of the sequence, and record the chromatogram.

[0021]

[0022]

[0023] Example 2 Limit of Quantitation and Limit of Detection Dilution / blank solution: 50% acetonitrile

[0024] After the system is stable, inject 1 injection of LOD solution and each LOQ test solution for analysis. If the S / N meets the requirements, repeat the injection 6 times and record the chromatogram.

[0025]

[0026] Example 3 Accuracy experiment Solution preparation Diluent: 50% acetonitrile Blank solution: diluent Wiping fluid: 95% ethanol Reference solution: same as Example 1

[0027]

[0028] After the system is stable, inject 1-3 injections of blank solution, 5 injections of reference solution 1, 1 injection of reference solution 2, and 1 injection of each concentration test solution. In the sequence, inject reference solution 1 every 8 hours and at the end to return to calibration and record the chromatogram.

[0029]

[0030]

Claims

1. A method for detecting the residual content of olmesartan medoxomil, olmesartan and hydrochlorothiazide in a granulator, comprising the following steps: (1) Sampling: wipe the sampling site with a cotton swab, add the sample to the acetonitrile solution, and after ultrasonic treatment, take the supernatant to obtain the test solution; (2) Solution preparation: Reference solution: 50% acetonitrile solution with a concentration of 7.5 μg / mL of olmesartan medoxomil and 1.1 μg / mL of hydrochlorothiazide; Olmesartan test solution: 50% acetonitrile solution with a concentration of 7.0 μg / mL of olmesartan; phosphate buffer; (3) Chromatographic injection: A chromatographic column with octylsilane bonded silica as the filler was selected, with a solution of acetonitrile:phosphate buffer in a volume ratio of 20:80 as the mobile phase A and a solution of acetonitrile:phosphate buffer in a volume ratio of 80:20 as the mobile phase B, and linear gradient elution was performed.

2. The method according to claim 1, characterized in that In the step (1), the cotton swab is first added with ethanol for cleaning, and after being taken out and shaken until there are no droplets, the sampling area is wiped with the cotton swab for sampling, and the sampling area size is 25 square centimeters. Subsequently, the cotton swab head is placed in a test tube, 10.0 mL of 50% acetonitrile solution is added, ultrasonic treatment is performed for 10 minutes, centrifuged, and the supernatant is taken to obtain the test solution.

3. The method according to claim 1, characterized in that The phosphate buffer in step (2) is a 1.36 g / L potassium dihydrogen phosphate solution, and the pH value is adjusted to 2.5 with phosphoric acid.

4. The method according to claim 1, characterized in that During the linear gradient elution, the flow rate was 1.5 ml per minute; the detection wavelength was 225 nm; the column temperature was 30° C.; the injector temperature was 5° C.; the injection volume was 10 μl, and the elution gradient was: 。

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