Detection reagent kit for pesticide residue, its preparing method and its use
A technology of molecules and monomers, applied in the field of pesticide residue detection kits
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Embodiment 1
[0035] The head of the sensitive housefly was frozen in liquid nitrogen and separated, and placed in a homogenizer at a ratio of 1:5 between the head of the housefly and the homogenate (pH 7.5, 0.1 mol / L). After the homogenate was subjected to differential centrifugation at 1000 g, 10000 g and 100000 g, the supernatant was precipitated with ammonium sulfate (50-60% saturation), and then desalted with a G25 column. After desalination, freeze-dry to make dry powder. Dissolve the dry powder in the buffer solution (pH7.5, 0.1mol / L) (w / v: 1:1) and pass it through the G200 chromatography column and the affinity chromatography column, collect the active eluate and freeze-dry to obtain the dry powder, namely It is acetylcholinesterase enzyme powder. Mix enzyme powder and protective agent (sucrose: cyclodextrin: mannitol = 1:4:5) at a ratio of 1:100 (w / w), and after solidification, it will be the enzyme source (A) of the detection kit; follow the above method Substrate and chromogeni...
Embodiment 2
[0037]The head of the sensitive housefly was frozen in liquid nitrogen and separated, and placed in a homogenizer at a ratio of 1:5 between the head of the housefly and the homogenate (pH 7.5, 0.1 mol / L). After the homogenate was subjected to differential centrifugation at 1000g, 10000g and 100000g, the supernatant was precipitated with ammonium sulfate (50-60 saturation), and then desalted with a G25 column. After desalination, it is freeze-dried to make dry powder. Dissolve the dry powder in distilled water (w / v: 1:1), centrifuge through the sucrose density gradient, collect the components of the 10%-20% sucrose gradient section, pass through the G200 chromatography column, collect the active eluent and freeze-dry it to obtain Dry powder is acetylcholinesterase enzyme powder. Mix the enzyme powder and the protective agent (starch: cyclodextrin: mannitol = 1:5:5) at a ratio of 1:150 (w / w), and after curing, it will be the enzyme source (A) of the detection kit; follow the ab...
Embodiment 3
[0056] The kit prepared according to Example 1 was stored at 20-30° C. in the dark for three weeks, and the enzyme activity at each time point was measured by conventional methods. The unit of enzyme activity value is ΔOD412 value / 3 minutes
[0057] The impact of table 3 room temperature preservation on the activity of acetylcholinesterase (in the table is the average value of 3 times of determination)
[0058] time (days)
[0059] It can be seen from the above table 3 that the acetylcholinesterase in the kit of the present invention can be stored at room temperature for three weeks without affecting its enzyme activity, therefore, the detection of pesticide residues can be carried out at room temperature.
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