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Method for detecting fluoxetine and norfluoxetine

A norfluoxetine and detection method technology, which is applied in the detection field of fluoxetine and norfluoxetine, and can solve the problems of long sample detection time, many organic solvents, and time-consuming

Pending Publication Date: 2021-03-30
BEIJING HARMONY HEALTH MEDICAL DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the detection of fluoxetine and norfluoxetine by existing high-performance liquid chromatography usually requires more complex pretreatment of the sample to be tested, many organic solvents used, more time-consuming, and requires long-term chromatographic separation and Cleaning and re-equilibration process resulting in longer sample testing times

Method used

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  • Method for detecting fluoxetine and norfluoxetine
  • Method for detecting fluoxetine and norfluoxetine
  • Method for detecting fluoxetine and norfluoxetine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] Embodiment 1: prepare the standard solution of series concentration

[0104] (a) Preparation of standard working solution

[0105] Accurately weigh 0.5 mg of fluoxetine standard substance and place it in a 5 mL volumetric flask, dissolve it with methanol solution, and set the volume to the mark line of the volumetric flask to obtain a standard stock solution A of 100 μg / mL.

[0106] Accurately weigh 0.5 mg of norfluoxetine standard substance and place it in a 5 mL volumetric flask, dissolve it with methanol solution, and set the volume to the mark line of the volumetric flask to obtain a standard stock solution B of 100 μg / mL.

[0107] Mix standard stock solution A and standard stock solution B, dilute with aqueous solution containing 70% methanol as the diluent to obtain a series of standard working solutions containing fluoxetine and norfluoxetine with different concentrations, and store them at -80°C ;

[0108] Wherein, the concentrations of fluoxetine and norfluox...

Embodiment 2

[0115] Embodiment 2: fitting standard curve equation

[0116] The standard solutions of seven concentrations in Example 1 were detected by liquid chromatography, and the chromatograms of standard solutions of seven different concentrations of fluoxetine and norfluoxetine were obtained.

[0117] Obtain respectively the chromatographic peak area of ​​fluoxetine, the chromatographic peak area of ​​norfluoxetine and the chromatographic peak area of ​​internal standard in seven kinds of standard solutions from the standard solution chromatogram of above-mentioned fluoxetine and norfluoxetine, The ratio of the chromatographic peak area of ​​the fluoxetine obtained in the chromatogram of the standard solution of the above-mentioned each concentration and the chromatographic peak area of ​​the internal standard is used as the ordinate y1 of the first standard curve equation, and the fluoxetine in the above-mentioned standard solution is The ratio of the concentration of Tin to the con...

Embodiment 3

[0128] Embodiment 3: the pretreatment of sample to be tested

[0129] 3.1 Take 2 mL of the sample to be processed, centrifuge at a speed of 3500 rpm for 10 min, take the supernatant serum or plasma as the first supernatant, and store the above serum or plasma at -20°C until analysis.

[0130] 3.2 Use a pipette gun to pipette 10 μL of the internal standard working solution in step (b) of Example 1 into a 1.5 mL centrifuge tube, then add 100 μL of the first supernatant in step 3.1, and then add 1000 μL of methyl tert-butyl Ether extractant, vortex mixing at 2500rpm for 10min, and high-speed centrifugation at 14000rpm for 5min, pipette 900 μL of the second supernatant (upper organic phase) after centrifugation, and use nitrogen gas to pipette the second supernatant Blow dry the supernatant, add 100 μL aqueous solution containing 70% methanol successively as complex solution, vortex mix at 2500 rpm for 1.0 min, and centrifuge at 14000 rpm for 5 min at high speed, take the third su...

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Abstract

The invention provides a method for detecting fluoxetine and norfluoxetine. The method comprises the following steps of: preparing standard solutions which have at least three kinds of concentrationsand contain fluoxetine, norfluoxetine and an internal standard substance; detecting each standard solution by utilizing a liquid chromatograph under a preset detection condition to obtain a first detection result; fitting a first standard curve equation of the fluoxetine and a second standard curve equation of the norfluoxetine according to the first detection result and the concentrations of thefluoxetine, norfluoxetine and internal standard substance in the standard solutions; carrying out centrifugal treatment on a to-be-treated sample, and taking first supernatant obtained after the centrifugal; adding the internal standard substance and an extraction agent into the first supernatant, carrying out vortex uniform mixing, and carrying out extraction to obtain the to-be-detected sample;detecting the to-be-detected sample by utilizing the liquid chromatograph under a detection condition to obtain a second detection result; and based on the first standard curve equation, the second standard curve equation and the second detection result, obtaining the concentrations of the fluoxetine and norfluoxetine in the to-be-detected sample. The method can shorten sample detection time.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a detection method for fluoxetine and norfluoxetine. Background technique [0002] Fluoxetine is a selective 5-HT reuptake inhibitor, white or off-white crystalline powder, easily soluble in methanol or ethanol, soluble in acetonitrile, acetone or chloroform, slightly soluble in ethyl acetate, dichloromethane or water. Norfluoxetine, also known as norfluoxetine, is a yellow solid. [0003] At present, the method usually used to detect the content of fluoxetine and norfluoxetine in a sample is high performance liquid chromatography. However, the detection of fluoxetine and norfluoxetine by existing high-performance liquid chromatography usually requires more complex pretreatment of the sample to be tested, many organic solvents used, more time-consuming, and requires long-term chromatographic separation and The cleaning and re-equilibration process results in longer...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/74
CPCG01N30/06G01N30/74
Inventor 赵金宝贾永娟倪君君
Owner BEIJING HARMONY HEALTH MEDICAL DIAGNOSTICS CO LTD
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