A method for detecting adrenaline substances in serum and its enrichment material
An epinephrine and detection method technology, applied in the field of analytical chemistry detection, can solve the problems of enrichment, large differences in processing methods, and no use, achieving good repeatability, overcoming interference and false positive problems, and improving the accuracy of results. Effect
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Embodiment 1
[0052] Embodiment 1: Test the accuracy comparison between different fillers.
[0053] Respectively use benzenesulfonic acid group bonded phase packing 1 bonded to the surface of silica gel with linear carbon chains, wherein the carbon chain length is 15, the particle size is 30um, and the pore size Specific surface area 300m 2 / g filler 2 with benzene rings bonded to the surface of silica gel.
[0054] Filler 1 structure.
[0055] Filler 2 structure.
[0056] Using the same processing method and detection conditions, take 100 mg of packing and 2 mL of solid phase column, add 1 mL of MeOH: H 2 After activation with O (10:90v / v), add 1mL MeOH:NH 4 FA:H 2 O (90:5:5, v / v / v) for equilibrium, and then after loading all the sample solution, add 500uLMeOH:NH 4 FA:H 2 O (90:5:5, v / v / v) and 500uL MeOH:H 2 O (10:90, v / v) rinse. Finally add 200uLMeOH: NH 4 FA:H 2 O: Ammonia water (90:2:7:1, v / v / v / v) was eluted, and all the eluent was collected and put into a sample injecti...
Embodiment 2
[0059] Embodiment 2: The detection limit comparison of different pretreatment methods.
[0060] Using the method of protein precipitation, take 200ul of serum sample, add 400uL of 0.2% formic acid acetonitrile solution to precipitate, put it in the refrigerator for 30min, centrifuge at 12000r / min for 3min to take the supernatant, and inject 10uL. Using the method of solid phase extraction, take 200uL serum into a 1.5mL EP tube, then add 20uL internal standard solution and vortex mix, then add 700μL methanol, vortex mix for 30 seconds, sonicate for 3min,; the linear alkyl group on the surface of the silica gel is connected The benzenesulfonic acid group bonded phase filler has a carbon chain length of 15, a particle size of 30um, and a pore size of Specific surface area 300m 2 / g.
[0061] packing structure
[0062] 500mg packing and 5mL solid phase column, add 3mL MeOH: H 2 After activation with O (10:90v / v), add 3mL of MeOH:NH 4 FA:H 2 O (90:5:5, v / v / v) to equilibrat...
Embodiment 3
[0065] Example 3: Detection of serum samples.
[0066] Take 200uL of serum, put it into a 1.5mL EP tube, add 20uL of internal standard extraction solution and vortex mix, then add 700μL of methanol, vortex mix for 30 seconds, and sonicate for 3min; the surface of the silica gel is bonded with benzenesulfonic acid groups linked by linear alkyl groups The group bonded phase filler has a carbon chain length of 15, a particle size of 30um, and a pore size of Specific surface area 300m 2 / g, 500mg packed in a 5mL solid-phase column, add 3mLMeOH:H 2 After activation with O (10:90v / v), add 3mL MeOH:NH 4 FA:H 2 O (90:5:5, v / v / v) to equilibrate, and then add 1 mL of MeOH:NH 4 FA:H 2 O (90:5:5, v / v / v) and 1 mL MeOH:H 2 O (10:90, v / v) rinse. Finally add 500uLMeOH: NH 4 FA:H 2 O: HCl (90:2:7:1, v / v / v / v) was eluted, and all eluents were collected, put into injection vials, entered into ultra-high performance liquid chromatography tandem mass spectrometry detection, and brought in...
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