Method for detecting arachidonic acid-like substances
A technology for the detection of arachidonic acid and its detection method, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of cumbersome and complicated preparation methods, high cost of pretreatment preparation methods, low sample content and difficulty, and achieve high extraction rate and improved detection The effect of improving sensitivity and signal-to-noise ratio
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Embodiment 1
[0028] Ultra-high performance liquid chromatography-triple quadrupole / linear ion trap tandem mass spectrometry (UPLC-QTRAP) technology was adopted, the mass spectrometer was AB Sciex Q-TRAP 6500, and the analysis software was Multi Quant.
[0029] Take 100uL of human plasma sample in EP tube, add 300uL of water (containing 0.005% BHT);
[0030] Add 500uL of MTBE (methyl tert-butyl ether) to the EP tube, vortex and mix well, centrifuge at 10000rpm for 3min at 4°C, transfer the upper layer organically to a new EP tube, and keep the lower aqueous phase;
[0031] Add 500 uL of ethyl acetate to the aqueous phase of the lower layer, vortex and mix well, centrifuge at 10,000 rpm for 3 min at 4°C, and combine the upper organic phase with the upper organic phase obtained in the previous step;
[0032] After the combined upper organic phases were blown dry with nitrogen, they were reconstituted with acetonitrile / isopropanol (1:1), filled into sample vials, and tested on the machine.
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Embodiment 2
[0049] Embodiment 2 (comparative example):
[0050] Take 100uL of plasma sample in EP tube, add 300uL of water (containing 0.005% BHT);
[0051] Add 500uL of MTBE (methyl tert-butyl ether) to the EP tube, vortex and mix well, centrifuge at 10000rpm for 3min at 4°C, transfer the upper layer organically to a new EP tube, and keep the lower aqueous phase;
[0052] Add 500 uL of MTBE (methyl tert-butyl ether) to the lower aqueous phase, vortex and mix well, centrifuge at 10,000 rpm for 3 min at 4°C, and combine the upper organic phase with the upper organic phase obtained in the previous step;
[0053] After the combined upper organic phases were blown dry with nitrogen, they were reconstituted with acetonitrile / isopropanol (1:1), filled into sample bottles, and waited for detection on the machine.
Embodiment 3
[0054] Embodiment 3 (comparative example):
[0055] Take 100uL of plasma sample in EP tube, add 300uL of water (containing 0.005% BHT);
[0056] Add 500uL of ethyl acetate to the EP tube, vortex and mix well, centrifuge at 10000rpm for 3min at 4°C, transfer the upper layer organically to a new EP tube, and keep the lower aqueous phase;
[0057] Add 500 uL of ethyl acetate to the aqueous phase of the lower layer, vortex and mix well, centrifuge at 10,000 rpm for 3 min at 4°C, and combine the upper organic phase with the upper organic phase obtained in the previous step;
[0058] After the combined upper organic phases were blown dry with nitrogen, they were reconstituted with acetonitrile / isopropanol (1:1), filled into sample bottles, and waited for detection on the machine.
[0059] Examples 1-3 Extraction methods The comparison of signal-to-noise ratios for extracting arachidonic acid-like substances is shown in Table 1. Examples 1-3 Extraction methods The extraction rates ...
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