Immunomagnetic chemiluminescence detection kit for chlorpromazine and application thereof
A chemiluminescence detection and chemiluminescence technology, which is applied in the field of immunology detection, can solve the problems of low sensitivity, high false negative rate and false positive rate, and achieve high sensitivity and specificity, short detection time, and full automation
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[0030] Example 1: Preparation of Specific Components of Chlorpromazine Magnetic Immunochemiluminescent Detection Kit
[0031] 1. Synthesis of chlorpromazine hapten (synthetic route sees attached figure 1 ) and identification
[0032] A.1.0g of N1-(5-chloro-2-iodobenzene)-N3 compound is dissolved in anhydrous N,N-dimethylformamide (DMF), add 0.8g of cuprous iodide, add sodium hydride 0.3g, add 0.66g of 3-bromo-2-thiophenol, heat in an oil bath, react at 100°C for 4h, stop the reaction, cool to room temperature, add water, extract with ethyl acetate, evaporate to dryness by rotary evaporation, put on a silica gel column, petroleum ether / ethyl acetate (v / v, 10 / 1) was eluted and separated to obtain the intermediate hydroxychlorpromazine 0.9g, with a yield of 91.84%;
[0033] B. Take 0.9 g of hydroxychlorpromazine, add acetonitrile to dissolve and clarify, add 0.54 g of iodoacetaldehyde, add 0.43 g of potassium carbonate, stir at 50 ° C for 3 h, stop the reaction, add water, ext...
Example Embodiment
[0055] Example 2: Construction of Chlorpromazine Magnetic Immunochemiluminescence Detection Kit
[0056] Set up the chlorpromazine magnetic immunochemiluminescence detection kit so that it includes the following components:
[0057] (1) horseradish peroxidase-labeled chlorpromazine hapten;
[0058] (2) The enzyme-labeled antigen diluent is pH 7.2-7.6, Na 3 PO 4 A buffer solution with a concentration of 0.01mol / L and a NaCl concentration of 0.25mol / L;
[0059] (3) Chlorpromazine monoclonal antibody labeled with immunomagnetic beads;
[0060] (4) Magnetic label antibody diluent is pH 7.2-7.6, Na 3 PO 4 A buffer solution with a concentration of 0.01mol / L and a NaCl concentration of 0.25mol / L;
[0061] (5) The concentrations of the standard solution of chlorpromazine series are 0μg / L, 0.05μg / L, 0.15μg / L, 0.45μg / L, 1.35μg / L, 4.05μg / L;
[0062] (6) The chemiluminescence substrate A solution is trishydroxymethylaminomethane solution containing luminol and p-cresol, and the che...
Example Embodiment
[0065] Embodiment 3: the detection of residual amount of chlorpromazine in the sample
[0066] 1. Sample pretreatment
[0067] (1) Animal tissue (chicken, pork, fish, shrimp) samples: Weigh 2.0g±0.05g homogenized tissue samples into a 50mL polystyrene centrifuge tube, add 2mL 2.5% trichloroacetic acid and 6mL acetone, respectively, Shake with a shaker for 5 minutes, mix well, centrifuge at 3000g room temperature (20-25℃ / 68-77℉) for 5min; pipette 2mL of the supernatant into a 50mL polystyrene centrifuge tube, add 2mL of 2mol / L sodium hydroxide solution and 4.5mL of n-hexane, oscillate with a shaker for 5min, mix well, centrifuge at 3000g room temperature (20-25℃ / 68-77℉) for 5min; transfer 3mL of the upper organic phase to a 10mL clean and dry glass tube, store at 50-60℃ ( 122-140℉) in a water bath and blow dry under nitrogen flow; add 0.5mL complex solution and vortex for 2min with a vortexer to dissolve the dried residue; take 50μL for analysis.
[0068] (2) Milk sample: pip...
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