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A method, kit and application for measuring phenobarbital in blood

A technology based on phenobarbital and phenobarbital, which is applied in the field of blood drug concentration determination, can solve the problems of large amount of organic reagents used, less selection space, and large amount of biological testing materials, so as to reduce the influence of interfering substances and operate Simple and fast, eliminate the effect of matrix effect

Active Publication Date: 2020-09-08
NINGBO MUNICIPAL CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, liquid-liquid extraction and solid-phase extraction are more popular, but the liquid-liquid extraction uses a large amount of organic reagents and pollutes the environment. The solid-phase extraction used in the solid-phase extraction is small, the cost of the column is high, and the selection space is small.
The column switching method currently has a single packing material for the pretreatment column, and its use is limited.
[0004] There have been publicly reported liquid chromatography-mass spectrometry (LC / MS-MS) for the determination of barbiturates in blood. However, it has been proved in practice that the methods reported have low sensitivity. Due to the shortcomings of large amount of biological testing materials and long analysis time, it is difficult to meet the needs of sensitive, simple, fast and accurate analysis for the detection of such drugs in clinical and judicial appraisal

Method used

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  • A method, kit and application for measuring phenobarbital in blood
  • A method, kit and application for measuring phenobarbital in blood
  • A method, kit and application for measuring phenobarbital in blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The mensuration of phenobarbital content in the blood sample of embodiment 1

[0045] 1.1 Preparation of standard stock solution

[0046] Accurately draw 100 μL of phenobarbital standard stock solution (1.0 mg / L) into a 2 mL volumetric flask, dilute to volume with acetonitrile, and prepare a 50.0 μg / L standard stock solution.

[0047] 1.2 Preparation of standard solution series

[0048] Accurately draw the standard stock solution of phenobarbital (50.0 μg / L) and prepare the concentrations of 0.05 μg / L, 0.1 μg / L, 0.5 μg / L, 1.0 μg / L, 2.5 μg / L, 5.0 μg / L, 10.0μg / L, 25.0μg / L, 50.0μg / L standard solution series.

[0049] 1.3 Chromatographic conditions

[0050] Chromatographic column: Waters ACQUITY UPLC BEH C18 column (100mm×2.1mm, 1.7μm); flow rate: 300μL / min; column temperature: 40℃; mobile phase: acetonitrile-0.1% formic acid aqueous solution (20:80, V / V); Injection volume: 5 μL.

[0051] 1.4 Mass Spectrometry Conditions

[0052] Ion source: electrospray ion source (E...

Embodiment 2

[0055] The selection of embodiment 2 chromatographic conditions

[0056] 2.1 Selection of chromatographic column

[0057] The present invention has tested 5 kinds of C18 chromatographic columns produced by different companies, respectively Waters Acquity UPLC BEHC18 column (100mm × 2.1mm, 1.7μm), Waters Xbridge C18 column (100mm × 2.1mm, 3.5μm), Phenomenex Kinetex C18 column (50mm ×2.1mm, 2.6μm), Shimadzu Shim-pack XR-ODSII column (150mm×2.0mm, 2.2μm) and Shimadzu Shim-pack XR-ODSIII column (150mm×2.0mm, 2.2μm) for phenobarbital drug separate effects. The results show that: SHIMADZU Shim-pack XR-ODSII column (150mm×2.0mm, 2.2μm) has the highest response value and better effect. Combined with the overall situation, Waters Acquity UPLC BEH C18 column (100mm×2.1mm, 1.7μm) was selected. The chromatograms of phenobarbital in 5 chromatographic columns are as follows figure 1 shown.

[0058] 2.2 Selection of mobile phase

[0059] The present invention has tested the impact of di...

Embodiment 3

[0064] The selection of embodiment 3 mass spectrometry conditions

[0065] 3.1 Selection of ion spray voltage

[0066] The present invention tests the responses of different ion spray voltages to various target compounds, including -1500, -2500, -3500 and -4500V. The results show that the ion spray voltage is the best when the voltage is -4500V, and the change of responsivity is as follows: Figure 5 shown.

[0067] 3.2 Selection of ion source temperature

[0068] The present invention tests the response of different ion source temperatures to each target compound, and mainly selects the following five ion source temperatures: 300, 400, 500, 600, and 700°C. The results show that the overall effect is better when the ion source temperature is 600°C.

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Abstract

The invention discloses a method, kit and application for measuring phenobarbital in blood, and belongs to the technical field of blood drug concentration determination. The method adopts HPLC-MS / MS to measure the content of phenobarbital in blood samples, and specifically includes the following Steps: (1) blood sample pretreatment; (2) preparation of standard working solution; (3) HPLC-MS / MS detection; the present invention adopts dispersive solid-phase extraction method, and uses PSA and PLS as dispersive solid-phase extraction agents to remove plasma The impurities in the plasma, anhydrous magnesium sulfate and anhydrous sodium sulfate remove the water in the plasma and reduce the influence of the matrix on the determination. A new method for the determination of phenobarbital in plasma samples by a green dispersive solid-phase extraction-liquid chromatography-tandem mass spectrometry method with satisfactory results.

Description

technical field [0001] The invention relates to the technical field of measuring blood drug concentration, in particular to a method, kit and application for measuring phenobarbital in blood. Background technique [0002] Sedative drugs are commonly used clinical drugs, which can inhibit the central nervous system and are mainly used for insomnia, anxiety, tension, fear, epilepsy and preoperative sedation. Sedative drugs are usually divided into benzodiazepines (estazolam, midazolam, alprazolam, triazolam, diazepam, clonazepam, lorazepam, nitrazepam, Xazepam, chlordiazepoxide, etc.), barbiturates (barbital, phenobarbital, amobarbital, etc.), phenothiazines (chlorpromazine, promethazine, etc.), quinazolones class (methaquinone, etc.) and imidazopyridines (zolpidem, etc.), etc. Research practices have shown that barbiturates are addictive, and their therapeutic dose is close to the toxic dose, so they have been gradually replaced by benzodiazepines in clinical practice. How...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/08G01N30/14G01N30/32G01N30/34G01N30/72
CPCG01N30/02G01N30/06G01N30/08G01N30/14G01N30/32G01N30/34G01N30/72G01N2030/047G01N2030/062G01N2030/146G01N2030/324
Inventor 金米聪陈晓红林梦周健俞建成
Owner NINGBO MUNICIPAL CENT FOR DISEASE CONTROL & PREVENTION
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