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Method for simultaneously detecting concentration of aromatase inhibitor, type 5 phosphodiesterase inhibitor and metabolites of aromatase inhibitor and type 5 phosphodiesterase inhibitor in human plasma

A type of phosphodiesterase and phosphodiesterase technology, applied in the field of clinical blood drug concentration monitoring, can solve the problems of narrow linear range, high detection cost, low sensitivity, etc., achieve good precision and accuracy, and extraction recovery rate High, high-sensitivity effects

Active Publication Date: 2021-07-23
金橄榄科技(上海)有限公司
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Problems solved by technology

[0007] It has been reported that liquid chromatography-tandem mass spectrometry is the most widely used method for the determination of AIs or PDE5is and their metabolites in human plasma, but these methods have more or less shortcomings: such as sample pretreatment is more complicated or Time-consuming, low sensitivity, long detection time, narrow linear range, large amount of plasma or organic reagents, etc., resulting in heavy workload, high detection cost, or failure to meet clinical needs; and the detection standards of these methods are inconsistent, so that they cannot be simultaneously Detect different drug concentrations in the same patient's plasma sample, which increases the workload and patient waiting time, and wastes a large amount of patient plasma

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  • Method for simultaneously detecting concentration of aromatase inhibitor, type 5 phosphodiesterase inhibitor and metabolites of aromatase inhibitor and type 5 phosphodiesterase inhibitor in human plasma
  • Method for simultaneously detecting concentration of aromatase inhibitor, type 5 phosphodiesterase inhibitor and metabolites of aromatase inhibitor and type 5 phosphodiesterase inhibitor in human plasma
  • Method for simultaneously detecting concentration of aromatase inhibitor, type 5 phosphodiesterase inhibitor and metabolites of aromatase inhibitor and type 5 phosphodiesterase inhibitor in human plasma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Example 1 Detection of Aromatase Inhibitors and Type 5 Phosphodiesterase Inhibitors and Their Metabolite Concentrations in Plasma

[0077] 1. Sample pretreatment

[0078] A stock solution of bifonazole was prepared with methanol as a solvent. In the stock solution of bifonazole, the concentration of bifonazole was 1 mg / mL, and it was sealed in a 4° C. refrigerator for future use.

[0079] The stock solution of bifonazole was diluted with methanol aqueous solution with a volume ratio of 1:1 to prepare an aqueous methanol solution of bifonazole, wherein the concentration of bifonazole was 0.25 μg / mL. The methanol aqueous solution of bifonazole was mixed with acetonitrile in a volume ratio of 1:20 to obtain a protein precipitant.

[0080] Take a human plasma sample, add 2 times the volume of protein precipitant, vortex for 3 min to precipitate protein, centrifuge at 12000g / min for 10 min at 4°C, take 100 μL of supernatant and put it in a sample injection tube to obtain a ...

Embodiment 2

[0100] Example 2 Quality control solution

[0101] The stock solutions of aromatase inhibitor, phosphodiesterase inhibitor type 5 and their metabolite standard products in steps 1 and 2 of Example 1 above and the stock solutions of bifonazole were diluted with an aqueous methanol solution with a volume ratio of 1:1. Stock solutions, respectively obtain quality control solutions of different concentrations of each component and internal standard, and mix the quality control solutions of different concentrations according to each concentration gradient to obtain a mixed quality control solution.

[0102] Take a series of 90 μL blank plasma samples, add 10 μL mixed quality control solution of a certain concentration gradient respectively, and use step 2 of the above Example 1 to process, and sequentially prepare quality control plasma solutions of different concentrations. The quality control plasma solutions are quantitative The lower limit LLOQ, the low quality control LQC, the...

Embodiment 3

[0105] Example 3 Methodological verification

[0106] Methodological validation of the method in the present invention mainly includes selectivity, linearity, precision and accuracy, matrix effect and recovery rate, and stability.

[0107] 1. Selective

[0108] 100 μL of human blank plasma sample was taken and processed according to step 1 of Example 1, wherein the methanol aqueous solution of bifonazole was not added to the protein precipitation agent. Take 100 μL of LLOQ plasma sample and process according to step 1 of Example 1. Then, the human blank plasma sample and the LLOQ plasma sample were tested according to step 3 of Example 1. It can be found that anastrozole, letrozole, sildenafil, N-desmethylsildenafil, tadalafil and bifonazole have good peak shapes under the above chromatographic conditions. For blank plasma, the peak area of ​​the interference signal in blank plasma was compared with the peak area of ​​analyte and internal standard in LLOQ.

[0109] 2. Line...

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Abstract

The invention provides a method for simultaneously a detecting concentration of aromatase inhibitor, a type 5 phosphodiesterase inhibitor and the metabolites of the aromatase inhibitor and the type 5 phosphodiesterase inhibitor in human plasma. The method comprises the following steps of: adding a protein precipitant into a human plasma sample to precipitate protein in the human plasma sample; measuring the obtained sample solution by high performance liquid chromatography-mass spectrometry; determining the aromatase inhibitor, the 5 type phosphodiesterase inhibitor and the metabolites thereof in the sample solution according to retention time; and performing quantifying by adopting an internal standard curve method so as to determine the concentrations of the aromatase inhibitor, the 5 type phosphodiesterase inhibitor and the metabolites thereof in the sample solution. The method not only is simple and rapid and accurate in quantification, but also has the advantages of high sensitivity, high selectivity, good repeatability, high recovery rate and the like, and meets the clinical large-batch biological sample analysis requirements of simple operation, reliable data and controllable conditions.

Description

technical field [0001] The invention belongs to the technical field of clinical blood drug concentration monitoring, relates to a method for simultaneously detecting the drug concentrations of aromatase inhibitors and type 5 phosphodiesterase inhibitors in human plasma, and particularly relates to a method for simultaneously detecting aromatase inhibitors in human plasma High Performance Liquid Chromatography-Mass Spectrometry (HPLC-MS / MS) Method for Enzyme Inhibitors, Phosphodiesterase Type 5 Inhibitors and Their Metabolites Concentrations. Background technique [0002] The World Health Organization (WHO) stipulates that if the husband and wife do not use any contraceptive measures to live together for more than 1 year, and the woman is infertile due to the male factor, it is called male infertility. Male infertility is a worldwide problem, and its treatment methods are limited, especially because there are no clear and effective drugs. In recent years, clinicians have use...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/34G01N30/36G01N30/72
CPCG01N30/02G01N30/06G01N30/34G01N30/36G01N30/72G01N2030/045
Inventor 范国荣
Owner 金橄榄科技(上海)有限公司
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