Morphine detection method based on SPR technique
A detection method and morphine technology, which are used in measuring devices, material analysis by optical means, instruments, etc., to achieve the effect of convenient purchase and easy operation
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Embodiment 1
[0041] (1) Turn on the SPR instrument, unlock the SPR chip slot, place the SPR chip in the slot, and lock the chip slot; the temperature of the SPR instrument is set to 25°C;
[0042] (2) At a flow rate of 20 μL / min, inject 250 μL of a mixture of EDC and NHS in an equal volume ratio on the surface of the SPR chip to activate the carboxyl groups on the surface of the chip; the concentration of EDC is 0.4M, and the concentration of NHS is 0.1M;
[0043] (3) BSA-Morphine solution of 1 mg / mL pH5.0 is passed through the SPR chip after activating the surface carboxyl group at a flow rate of 20 μL / min, so that it is coupled to the activated carboxyl group, and the target response value (RU ) reaches 3000;
[0044] (4) Pass 100 μL of 1M ethanolamine hydrochloride solution at a flow rate of 20 μL / min on the SPR chip coupled with BSA-Morphine to block unreacted activated carboxyl sites;
[0045] (5) 40 μL of morphine antibody with gradient concentrations of 3.13 μg / mL, 6.25 μg / mL, 12.5...
Embodiment 2
[0053] (1) Turn on the SPR instrument, unlock the SPR chip slot, place the SPR chip in the slot, and lock the chip slot; the temperature of the SPR instrument is set to 25°C;
[0054] (2) Pass a mixture of 100 μL 0.2M EDC and 100 μL 0.1M NHS to the surface of the SPR chip at a flow rate of about 10 μL / min to activate the carboxyl groups on the surface of the chip;
[0055] (3) Pass 0.5mg / mLBSA-Morphine solution (pH value is 4.5) with the flow rate of 10μL / min on the SPR chip after the surface carboxyl group is activated in step (2), make it couple to the activated carboxyl group, require The target response value (RU) reached about 2000;
[0056] (4) Pass 75 μL of 2M ethanolamine hydrochloride solution at a flow rate of 10 μL / min on the SPR chip after BSA-Morphine coupling in step (3) to block unreacted activated carboxyl sites;
[0057] (5) At a flow rate of 10 μL / min, 30 μL of morphine antibody with gradient concentrations of 3.13 μg / mL, 6.25 μg / mL, 12.5 μg / mL, 25 μg / mL, 50...
Embodiment 3
[0065] (1) Turn on the SPR instrument, unlock the SPR chip slot, place the SPR chip in the slot, and lock the chip slot; set the temperature of the SPR instrument to 30°C;
[0066] (2) Pass a mixture of 150 μL 0.2M EDC and 150 μL 0.2M NHS to the surface of the SPR chip at a flow rate of about 30 μL / min to activate the carboxyl groups on the surface of the chip;
[0067] (3) Pass 2mg / mL BSA-Morphine solution (pH value is 5.5) with the flow velocity of 30 μ L / min on the SPR chip after the surface carboxyl group is activated in step (2), make it couple to the carboxyl group after activation, require The target response value (RU) reached about 3000;
[0068] (4) Pass 100 μL of 2M ethanolamine hydrochloride solution at a flow rate of 30 μL / min on the SPR chip after BSA-Morphine coupling in step (3) to block unreacted activated carboxyl sites;
[0069] (5) At a flow rate of 30 μL / min, 50 μL of morphine antibodies with gradient concentrations of 3.13 μg / mL, 6.25 μg / mL, 12.5 μg / mL, ...
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