Quick detection paper card for pesticide residue in vegetable and preparation method therof
A pesticide residue and vegetable technology, which is applied in the field of vegetable pesticide residue quick test paper card and its preparation field
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Embodiment 1
[0035] Embodiment 1, the screening of enzyme source
[0036] Using horse serum, human serum, bovine liver, bee head, housefly head and other animal organs or serum as materials, extract and screen the highly sensitive enzyme source-acetylcholinesterase according to conventional methods. Mash frozen horse serum, human serum, bovine liver, bee brain, and housefly brain respectively, add distilled water at a ratio of 1:9 (W / V), homogenate for 3 minutes, and centrifuge at 2000-3000r / min for 10 minutes , the supernatant was filtered through filter paper, freeze-dried, divided into equipment, and stored at -20°C. It was the enzyme source.
[0037] 1) Reagent preparation:
[0038] Enzyme liquid preparation: Dilute freeze-dried horse serum enzyme powder, serum enzyme powder, bovine liver enzyme powder, bee brain enzyme powder, and housefly brain enzyme powder with PH8. .
[0039] Pesticide concentration: 0.8ppm for methamidophos and 0.002ppm for carbofuran.
[0040] Substrate: Thi...
Embodiment 2
[0047] Embodiment 2: the selection of enzyme concentration
[0048] 1) Enzyme preparation: Dilute the freeze-dried human serum enzyme powder of the present invention with pH 8.0 and 0.1M phosphate buffer to 0.5, 0.75, 1.0, 1.25, and 1.5 units respectively. Add one drop (about 30 μl) onto a qualitative filter paper with a diameter of 1.0 cm to make an enzyme tablet.
[0049] 2) Substrate: Mix 0.0448g potassium ferricyanide (red blood salt) and 0.0715g potassium ferrocyanide (yellow blood salt), add 100ml distilled water (PH5.5) after mixing, add one drop (about 30μl) in side becomes 1.0cm 2 On the qualitative filter paper, add dropwise 30μl indole acetate 180mg / 10ml acetone solution to make a substrate sheet.
[0050] 3) The concentration of methamidophos is 0.1 mg / μl.
[0051] 4) Use distilled water for comparison
[0052] The results of the reaction of five kinds of enzyme concentration enzyme tablets with the pesticide methamidophos are shown in Table 3
[0053] Table 5...
Embodiment 3
[0060] Embodiment 3: the selection of optimal substrate concentration
[0061] 1) Preparation of substrate sheet: Mix 0.0448g potassium ferricyanide (red blood salt) and 0.0715g potassium ferrocyanide (yellow blood salt) into 100ml distilled water, take a drop of the mixture (about 30μl pH5.5) first Add dropwise on 3 pieces with a side length of 1.0cm 2 Qualitative filter paper, and then put a drop (about 30μl) of indole acetate 180mg / 10ml acetone solution, indole acetate 270mg / 10ml acetone solution, indole acetate 360mg / 10ml acetone solution on three pieces of qualitative filter paper On the surface, dry at room temperature for use as a substrate sheet.
[0062] 2) Preparation of enzyme tablets: Dilute the frozen enzyme powder extracted from the serum with pH 8.0 and 0.1M phosphate buffer to 0.5 and 0.75 units respectively. Add it dropwise on a qualitative filter paper with a diameter of 1.0 cm to make an enzyme tablet.
[0063] The results of reacting 0.5 units and 0.75 u...
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