Ferrohemoglobin adduct detecting method and application for evaluating in-body exposure of glycidol and glycidyl ester
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.
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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. Prepare a serie...
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