Device and method for determining 8-isoprostaglandin F2 alpha in urine

A prostaglandin and assay method technology, applied in the field of assay devices and devices for measuring 8-isomeric prostaglandin F2α in urine, can solve the problems of many factors of analysis error, complex background of urine sample matrix, troublesome operation and the like

Active Publication Date: 2020-03-24
CHINA TOBACCO YUNNAN IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the low content of this compound in urine samples and the presence of other compounds with similar structures, the sample usually requires solid-phase extraction or a combination of liquid-liquid extraction-solid-phase extraction for purification, concentration and quantification; sample pretreatment The process requires multiple transfers, which is not only troublesome to operate, but also introduces many factors of analysis errors
On the other hand, the matrix background of urine samples is complex, and when HPLC / MS / MS is used for analysis, the matrix effect of the sample will also affect the analysis results

Method used

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  • Device and method for determining 8-isoprostaglandin F2 alpha in urine
  • Device and method for determining 8-isoprostaglandin F2 alpha in urine
  • Device and method for determining 8-isoprostaglandin F2 alpha in urine

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Experimental program
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Effect test

Embodiment 1

[0068] Such as figure 1 As shown, the device for measuring 8-hydroxyisomeric prostaglandin F2α in urine of this embodiment includes a six-way valve 7 with a sample inlet, a sampler 6, an enrichment pump 5, and an analysis pump 2 , a detector 4, an online solid phase extraction column 1, an analysis column 3 and a chromatographic workstation. The six-way valve 7 with the sample inlet is provided with seven ports, namely the first port to the seventh port.

[0069] The detector 4 is connected to one end of the analytical column 3, the other end of the analytical column 3 is connected to the third port, the analytical pump 2 is connected to the second port, the enrichment pump 5 is connected to the sampler 6, and the sampler 6 is connected to the seventh port .

[0070] The six-way valve 7 with a sample inlet is used for switching between the solid phase extraction column 1 and the analysis column 3; the solid phase extraction material of the solid phase extraction is graphene ...

Embodiment 2

[0079] The device of this embodiment is the same as that of Embodiment 1.

[0080] The present embodiment is based on the assay method of 8-isomer prostaglandin F2α in the urine of above-mentioned device, carries out as follows:

[0081] The test samples were urine samples from light smokers. Take 7.5 mL of smoker urine sample collected, add 200 µL of internal standard working solution, and dilute to 10 mL with methanol; shake well, bring back to the laboratory within 20 h, and centrifuge at 20,000 r / min for 10 min at high speed; Take about 5.0 mL of the supernatant, filter it with a 0.25 µm syringe filter, and use it for online solid-phase extraction enrichment.

[0082] The sample injection volume was 2.0 mL, and after the sample injection, the enrichment pump used 25% methanol as the mobile phase at a flow rate of 1.0 mL / min to transport through the solid phase extraction column for enrichment. After the sample completely passes through the solid-phase extraction column, ...

Embodiment 3

[0085] The device of this embodiment is the same as that of Embodiment 1.

[0086] The present embodiment is based on the assay method of 8-isomer prostaglandin F2α in the urine of above-mentioned device, carries out as follows:

[0087] The samples tested were urine samples from moderate smokers. Take 7.5 mL of smoker urine sample collected, add 200 µL of internal standard working solution, and dilute to 10 mL with methanol; shake well, bring back to the laboratory within 20 h, and centrifuge at 20,000 r / min for 10 min at high speed; Take about 5.0 mL of the supernatant, filter it with a 0.25 µm syringe filter, and use it for online solid-phase extraction enrichment.

[0088] The sample injection volume was 1.5 mL, and after the sample injection, the enrichment pump used 25% methanol as the mobile phase at a flow rate of 1.0 mL / min to transport through the solid phase extraction column for enrichment. After the sample has completely passed through the solid-phase extraction...

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Abstract

The invention relates to a device and a method for determining 8-isoprostaglandin F2 alpha in urine, and belongs to the field of analytical chemistry. The method comprises the following steps: injecting a prepared sample, conveying the sample by using an enrichment pump, and enriching the sample by using a graphene oxide bonded silica gel solid-phase extraction column; after the sample enrichmentis finished, switching through a six-way valve, conveying a mobile phase by using an analysis pump, eluting the solid-phase extraction column from a flow direction opposite to the sample enrichment direction, separating eluted components through the analysis column, and detecting and determining the content of to-be-analyzed components in the sample through a detector. By using the online solid-phase extraction method, high-multiple enrichment of the sample on the enrichment column is realized; under the precondition that a chromatographic peak is not obviously widened, the sample injection amount of a sample is increased by nearly three orders of magnitudes, the analysis sensitivity of the method is greatly improved, and the content of 8-isoprostaglandin F2 alpha with the order of magnitude of pg / mL in a urine sample can be accurately determined. And a sensitive, accurate and reliable new method is provided for research on correlation between smoking and oxidative stress reaction.

Description

technical field [0001] The invention relates to a measuring device, especially a device for measuring 8-isomeric prostaglandin F2α in urine, and also relates to a measuring method, which belongs to the field of analytical instruments. Background technique [0002] Oxidative stress refers to the imbalance between oxidation and anti-oxidation in the body. Free radicals produce a negative effect in the body that tends to oxidize, which will lead to inflammatory infiltration of neutrophils, increased secretion of proteases, and the production of a large number of oxidized intermediates. In living organisms, free radicals and other highly reactive chemicals are difficult to measure directly due to their low concentrations and short lifetimes. Therefore, detection methods such as free radicals, active substances, oxidative stress, spin trapping, DNA markers, lipid and protein oxidative stress, cellular fluorescent probes, etc. may be possible in the qualitative detection of active...

Claims

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Application Information

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IPC IPC(8): G01N30/08G01N30/88
CPCG01N30/08G01N30/88
Inventor 李晶黄海涛杨叶昆刘欣孔维松高茜许永王晋米其利向海英曾婉利杨光宇李雪梅汪伟光
Owner CHINA TOBACCO YUNNAN IND
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