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Biological analysis method for concentration of tenofovir alafenamide and metabolite tenofovir in antiviral drug clinical research plasma sample

A technology for tenofovir alafenamide and antiviral drugs, which is applied in the directions of analysis materials, material separation, measuring devices, etc., to achieve the effects of simple pretreatment operation and easy storage.

Pending Publication Date: 2021-05-25
苏州海科医药技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are few technical reports on the simultaneous analysis of tenofovir alafenamide and tenofovir in plasma

Method used

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  • Biological analysis method for concentration of tenofovir alafenamide and metabolite tenofovir in antiviral drug clinical research plasma sample
  • Biological analysis method for concentration of tenofovir alafenamide and metabolite tenofovir in antiviral drug clinical research plasma sample
  • Biological analysis method for concentration of tenofovir alafenamide and metabolite tenofovir in antiviral drug clinical research plasma sample

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Experimental program
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Embodiment

[0030] pre-processing

[0031] The amount of plasma used in the present invention is only 100 μ L, which is suitable for clinical research and biological analysis. The extraction method in the present invention is selected from the protein precipitation method. Nofovir has a high recovery rate, and has simple operation, short extraction time, and no time-consuming concentration steps. Used with 96-well plates, it is suitable for high-throughput sample pretreatment in clinical research.

[0032] The specific preprocessing steps are as follows:

[0033] 1. Add 100 μL plasma sample, 50.0 μL internal standard solution (20.0 / 3.00 ng / mL), 300 μL methanol:acetonitrile (50:50, v / v) into the 96-well plate;

[0034] 2. Vortex to mix, centrifuge for 10min (4°C, 3900rpm);

[0035] 3. Transfer 100 μL supernatant to another clean 96-well plate;

[0036] 4. Add 300 μL aqueous solution containing 0.1% formic acid to dilute.

[0037] 5. The injection volume is 20.0 μL.

[0038] mass spec...

Embodiment 1

[0043] Abbreviation

[0044]

[0045] 1 material

[0046] 1.1 Instrument

[0047] Shimadzu Corporation LC-30AD Ultra High Performance Liquid Chromatography System and AB Sciex Triple Quad TM 6500 + A triple quadrupole tandem mass spectrometer equipped with an ESI source.

[0048] Analyst 1.6.3 software was used for data processing.

[0049] Beijing Sartorius Instrument Co., Ltd. CP225D analytical balance.

[0050] Japan Homer company Z326K desktop centrifuge

[0051] 5810R multifunctional desktop centrifuge, Eppendorf, Germany

[0052] 1.2 Reference substances and reagents

[0053] Tenofovir alafenamide (purity 98%) was purchased from TRC company; Tenofovir alafenamide (purity 99.7%, content 93.4%) was purchased from TLC company; Tenofovir alafenamide-d5 (purity 98% ) was purchased from TRC Company; tenofovir-d7 (chemical purity 99.8%, isotopic purity 99.6%) was purchased from TLC Company. Methanol (HPLC grade) and acetonitrile (HPLC grade) were purchased from Sig...

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Abstract

The invention relates to a biological analysis method for clinically researching the concentration of tenofovir alafenamide and metabolite tenofovir in a plasma sample through antiviral drugs, which is characterized in that the tenofovir alafenamide and metabolite tenofovir in plasma are analyzed through LC-MS / MS, and a protein precipitation pretreatment method is adopted; tenofovir alafenamide -d5 and tenofovir -d7 are taken as internal standards, an Eclipse Plus C18 chromatographic column is adopted, gradient elution is performed, and an electrospray ionization (ESI) source is used for tandem mass spectrometry detection. The method is easy and convenient to operate, high in analysis speed and high in sensitivity, and meets the requirement for large-batch sample analysis in clinical research.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a biological analysis method for the concentration of tenofovir alafenamide and metabolite tenofovir in plasma samples of antiviral drug clinical research. Background technique [0002] Tenofovir alafenamide is the prodrug of tenofovir, it is a new type of nucleotide reverse transferase inhibitor, through entering the DNA reverse transferase of hepatitis B virus, inhibiting the replication of DNA and used for Treatment of chronic hepatitis B. Tenofovir alafenamide fumarate tablets were developed by Gilead in the United States, and were approved by the FDA in October 2016 for the treatment of chronic hepatitis B in adults. The trade name is Vemlidy. Tenofovir alafenamide, as a lipophilic drug, is hydrolyzed to tenofovir by carboxylesterase in the human body. At present, there are few technical reports on the simultaneous analysis of tenofovir alafenamide and teno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/72G01N30/02G01N30/06
CPCG01N30/72G01N30/02G01N30/06
Inventor 陈云辉钟勘肖涛杨勇史中杰温凤娇刘旭凌周林芳姜金方周茂金
Owner 苏州海科医药技术有限公司
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