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Hemoglobin adduct assay method and application for evaluating glycidol and its esters exposure in vivo

A technology of hemoglobin and glycidol, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of low derivation efficiency and purification efficiency, unfavorable detection and detection of large batches of biological samples, lengthy processing operations, etc.

Active Publication Date: 2021-02-26
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The above-mentioned methods involve long pretreatment operations, low derivation efficiency and purification efficiency, and the detection sensitivity of the gas chromatography method is very limited, which is not conducive to the detection of large quantities of biological samples and the detection of samples with low dietary exposure levels

Method used

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  • Hemoglobin adduct assay method and application for evaluating glycidol and its esters exposure in vivo
  • Hemoglobin adduct assay method and application for evaluating glycidol and its esters exposure in vivo
  • Hemoglobin adduct assay method and application for evaluating glycidol and its esters exposure in vivo

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Effect test

Embodiment 1

[0097] Embodiment 1. A method for synchronous detection of hemoglobin adducts for evaluating dietary glycidol and its esters for medium and long-term exposure in vivo. The detection object is rat blood, and the following steps are carried out in sequence:

[0098] 1), preparation of dry hemoglobin powder:

[0099] Centrifuge 5ml of the whole blood sample in the anticoagulant tube at 3500rpm for 5min, remove the supernatant (plasma) and the middle layer (white membrane), and add 3ml of PBS solution or normal saline to the red blood cells left in the lower layer, and use Pipette the dropper evenly, suck off the supernatant, centrifuge at 3000rpm for 5min, repeat washing 3 times, and obtain 2ml of red blood cells.

[0100] Take 1 mL of red blood cells, transfer them to a centrifuge tube, add 5 mL of pure water to dilute the red blood cells 6 times, vortex and shake for 10 min, and put them into a -80°C refrigerator for quick freezing for 2 h. Afterwards, hemolysate was obtained ...

Embodiment 2

[0123] Embodiment 2. A method for synchronous detection of hemoglobin adducts for evaluating dietary glycidol and its esters for medium and long-term exposure in vivo. The detection object is human blood, and the following steps are performed in sequence:

[0124] 1), preparation of dry hemoglobin powder:

[0125] The blood of the crowd was collected in anticoagulant tubes, and the red blood cell pretreatment and dry hemoglobin powder were prepared. The processing method was the same as step 1) of Example 1.

[0126] 2), derivation and purification:

[0127] In 20 μL of mixed isotope internal standard, the concentration of d5-Gly-Val-PTH is 1 μg / mL, and the rest is the same as step 2) of Example 1.

[0128] 3) Chromatographic conditions

[0129] Identical to step 3) of embodiment 1.

[0130] 4) Mass spectrometry conditions

[0131] Identical to step 4) of embodiment 1.

[0132] 5) Results

[0133] The samples were quantified by the standard curve method. Configure a ser...

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Abstract

The invention discloses a hemoglobin adduct detection method for evaluating the exposure of glycidol and its esters in vivo, which comprises preparing hemoglobin dry powder, derivatizing and purifying the hemoglobin powder; Quantitative detection by mass spectrometry: using the liquid phase-mass spectrometry method above, the sample is quantitatively analyzed by the standard curve method. The present invention adopts isotope dilution ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS / MS) to quantify the hemoglobin adduct of glycidol, the analysis time is greatly shortened, and the Gly-Val-PTH spatial isomer is obtained at the same time Better separation enables simultaneous detection, which enables a more comprehensive assessment of the internal exposure of glycidol.

Description

technical field [0001] The invention relates to a method for synchronous detection of hemoglobin adducts for evaluating medium and long-term exposure of dietary glycidol and its esters in vivo and its application, belonging to the field of food safety. Background technique [0002] In 2009, the German Institute for Risk Assessment assessed the contamination of glycidyl esters in infant milk powder and edible oils (assuming that glycidyl esters are 100% converted to glycidol in the body), based on the genotoxicity and carcinogenicity of glycidol BMDL10 , the Margin of Exposure (MOE) ratio of infants and young children ingesting glycidyl esters in milk powder is 670, and the health risk is extremely high. Focusing only on 3-monochloro-1,2-diol (3-monochloropropane-1,2-diol; 3-MCPD) or 3-MCPD contamination and exposure will underestimate the health risks of these compounds. Therefore, all kinds of 3-MCPD Contamination of alcohol esters and glycidyl esters in food should also c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/60G01N30/86
CPCG01N30/02G01N30/06G01N30/60G01N30/8606G01N2030/027G01N2030/062
Inventor 章宇贾伟陈信宇
Owner ZHEJIANG UNIV
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