Time-resolved immunoassay kit for detecting chlorpromazine residues and detection method thereof
A detection kit and immunoassay technology, applied in the field of immunoassay, can solve the problems of lack of chlorpromazine time-resolved immunoassay kit, etc., and achieve the effect of simple sample pretreatment, short operation process and good effect
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Embodiment 1
[0044] A time-resolved immunoassay detection kit for detecting chlorpromazine residues, comprising a microtiter plate coated with chlorpromazine antigen, chlorpromazine antibody, lanthanide-labeled goat anti-rabbit or goat anti-mouse antibody (secondary antibody) ; the lanthanides include Eu 3+ , Tb 3+ 、Dy 3+ 、Sm 3+ and other rare earth elements.
[0045] The chlorpromazine antigen is obtained by coupling the chlorpromazine hapten with the carrier protein by carbodiimide method (EDC), active ester method (DCC, NHS), mixed anhydride method (isobutyl chloroformate). The carrier protein includes bovine serum albumin (BSA), human serum albumin (HSA), keyhole limpet hemocyanin (KLH) and other carrier proteins.
[0046] The invention provides a preparation method of the chlorpromazine hapten, which is to dissolve chlorpromazine in acetic anhydride, drop freshly prepared acetyl nitrate in an ice bath, and react to obtain CPZ-NO 2 , and then hydrogen reduction under the catalysis...
Embodiment 2
[0080] The present invention provides the method for detecting chlorpromazine residue simultaneously, comprises the following steps:
[0081] (1) Sample pretreatment;
[0082] a. Urine sample processing
[0083] Clear urine samples can be directly analyzed. If the urine sample is turbid, centrifuge at 2000r / min for 5min or filter, and use the supernatant for detection.
[0084] b. Feed sample processing
[0085] Crush the feed, weigh 2 g and place it in a 50 mL test tube, add 12 mL of 10% ammoniated methanol (9 parts of ammonia solution with a pH value of 9 to 10 + 1 part of methanol), mix well and shake for 1 to 2 minutes. Add 9 mL of ethyl acetate and shake for 20 min. Centrifuge at 5000r / min (4000g) for 10min. Take 500 μL of the organic phase in another test tube and dry it with nitrogen. Add 500 μL of sample diluent to dilute and detect directly. A dilution factor of 10 should be taken into account in the calculation of results.
[0086] c. Tissue (muscle, liver, k...
example 1
[0105] Hapten synthesis and identification
[0106] The synthesis of chlorpromazine hapten as figure 2 shown.
[0107] (1) Take by weighing 319mg (1mmol) chlorpromazine and dissolve in 5mL acetic anhydride, stir and dissolve fully in an ice bath;
[0108] (2) Get freshly prepared acetyl nitrate 4mmol and dissolve it in 1 mL of acetic anhydride and slowly drop it into the solution described in step (1), react in an ice bath for 3 hours, and after the reaction is completed, adjust the pH value of the product with ice-saturated sodium hydroxide solution To 8, the precipitated product is the nitrated compound of chlorpromazine, and the productive rate is 53%;
[0109] (3) Dissolve the nitrate of the above-mentioned chlorpromazine in 20mL ethanol, add 10% palladium carbon catalyst and obtain the target product under hydrogen reduction.
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