Method for detecting glimepiride

A detection method and detection condition technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of long analysis time of samples to be tested, long sample detection time, etc., and achieve the effect of shortening the sample detection time

Pending Publication Date: 2021-02-09
济南和合医学检验有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the existing detection methods usually use gradient elution, which needs to constantly change the concentration ratio of the elution mobile phase, and the analysis time of the sample to be tested is longer, resulting in a longer sample detection time.

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  • Method for detecting glimepiride
  • Method for detecting glimepiride
  • Method for detecting glimepiride

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0112] Embodiment 1: prepare the standard solution of series concentration

[0113] (a) Preparation of standard stock solution:

[0114] Accurately weigh 5 mg of glimepiride standard substance and place it in a 5 mL volumetric flask, dissolve it with methanol solution containing 50% acetonitrile, and set the volume to 5 mL to obtain a standard stock solution, and store it at -80°C.

[0115] (b) Preparation of standard working solution

[0116] Take an appropriate amount of the standard stock solution in step (a), dilute and mix it with an aqueous solution containing 90% methanol as a diluent to obtain a series of intermediate solutions containing 10-4000ng / mL glimepiride, and store them at -80°C save;

[0117] Among them, the standard working solution with different concentrations is a series of solutions containing glimepiride: 10ng / mL, 50ng / mL, 200ng / mL, 500ng / mL, 1000ng / mL, 2000ng / mL, 4000ng / mL.

[0118] (c) Preparation of internal standard stock solution

[0119] Take ...

Embodiment 2

[0124] Embodiment 2: fitting standard curve equation

[0125] The seven standard solutions in Example 1 were detected by liquid chromatography-mass spectrometer, and the chromatograms of seven standard solutions of glimepiride with different concentrations were obtained.

[0126] From the standard solution chromatogram of above-mentioned glimepiride, obtain respectively the corresponding peak area of ​​glimepiride and internal standard in seven kinds of standard solutions, obtain the glimepiride in the chromatogram of above-mentioned each kind of concentration standard solution The ratio of the peak area of ​​urea to the chromatographic peak area of ​​the internal standard is used as the ordinate y1 of the standard curve equation, and the ratio of the concentration in the above-mentioned glimepiride standard working solution to the concentration of the internal standard is used as the abscissa of the standard curve equation x1, perform linear regression on the data of differen...

Embodiment 3

[0140] Embodiment 3: the processing of test sample

[0141] 3.1 Take at least 500 μL of blood to be processed, centrifuge at a centrifugation speed of 3500 rpm for 10 minutes, take the supernatant serum or plasma as the first supernatant, and store the above serum or plasma at -20°C until analysis.

[0142]3.2 Use a pipette gun to pipette 10 μL of the internal standard working solution in Example 1 into each well of a 96-well plate, then add 50 μL of the serum or plasma in step 3.1, vortex and mix at 2000 rpm for 1 min, and then add the extractant After 500 μL of ethyl acetate solution, seal the 96-well plate (for example, seal the film with a plate sealer), vortex and mix at 2000 rpm for 3 minutes, and then centrifuge at 4000 rpm for 5 minutes. Supernatant liquid (upper organic phase) was 200 μL, and the pipetted second supernatant was blown dry with nitrogen gas, and 100 μL complex solution (aqueous solution containing 70% acetonitrile) was sequentially added, vortexed at 20...

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Abstract

The invention provides a glimepiride detection method which comprises the following steps: preparing standard solutions with at least three concentrations of glimepiride and internal standard substances, and ensuring that the quantities of the internal standard substances in the standard solutions are the same; detecting each standard solution by using a liquid chromatograph-mass spectrometer under a detection condition to obtain a first detection result corresponding to the standard solution; simulating a standard curve equation of qualified glimepiride according to each first detection result, the concentration of glimepiride in the standard solution and the concentration of an internal standard substance; taking a first supernatant of a sample to be treated after centrifugation; addingthe internal standard substance into the first supernatant, uniformly mixing in a vortex manner, sequentially adding an extracting agent, and extracting the first supernatant to obtain a sample to bedetected; detecting the sample to be detected by utilizing the liquid chromatograph-mass spectrometer under the detection condition to obtain a second detection result of the sample to be detected; and based on the standard curve equation and the second detection result, obtaining the concentration of glimepiride in the sample to be detected. The scheme can shorten the sample detection time.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a detection method of glimepiride. Background technique [0002] Glimepiride is a sulfonylurea compound, a white crystalline powder, odorless, tasteless, soluble in chloroform, slightly soluble in methanol or ethanol, insoluble in water or ether. [0003] At present, the method commonly used to detect the content of glimepiride in samples is liquid chromatography-mass chromatography. However, the existing detection methods usually use gradient elution, which needs to constantly change the concentration ratio of the elution mobile phase, and the analysis time of the sample to be tested is longer, resulting in a longer sample detection time. Contents of the invention [0004] The invention provides a detection method for glimepiride, which can shorten the sample detection time. [0005] In order to solve the above problems, the embodiment of the present invention p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/88
CPCG01N30/06G01N30/88
Inventor 许丽贾永娟倪君君
Owner 济南和合医学检验有限公司
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