Development Of Enzyme-linked Immunosorbent Assay For Chlorpromazine

Chlorpromazine hydrochloride (CPZ) belongs to the phenothiazine group of compounds, as a central dopamine receptor blocking agent, used to sedative anti-psychotic,suppress vomiting,tranquilizer hypothermia,reduced basal metabolism, it isα-adrengenergic receptor blocking agent,m-cholinergic blocking receptor agent , also affected incretion,etc,CPZ is used in human medicine as a tranquilliser, schizophrenia, hypothermic anesthesia, artificial hibernation.CPZ has been usually used as feed additive in animals with its special biological effects of calmness and hypnosis,However,CPZ has a toxic of leukocytopenia, postural hypotension,hepatosia ,tetter,contact dermatitis,tremor,cathisophobia, salivate, tardive dyskinesia,delayed cinesipathy,CPZ was ruled not t o be used in food animals by any way except for UK, in 1997 European union decree ban (EC)NO.17/97 :no chlorpromazine in feeding ,in 2002, China also decree ministry of agriculture number 176 and 235,no chlorpromazine in feeding and watering, Compare to physical and chemical inspection, immunology analysis has widely been application in drug residue ,it has many advantages, has more better sensitivity, rapidity, simplicity .The main contents and experimental results of this study were as follows:This study aimed at: the immunogenicity of CPZ, synthesis of complete antigen, preparation special properites of CPZ polyclonal antibody, two methods are synthesized by a chemical process. One method: Friedel-Crafts acylation reaction, and after alkalify, and introduction a active carboxyl in side ring then adopt EDC coupled BSA and OVA.Another method: azo-benzoic acid method , introduction a aromatic group, then used mixed anhydride coupled with BSA and OVA,and identification with UV, TOF-MS, EDC-BSA-CPZ was synthesized successfully and its conjugation ratio of CPZ to BSA was about5:1,mixed anhydride-CPZ-BSA was 15:1.Ten newzealand white rabbits were immunized with EDC-BSA-CPZ, mixed anhydride-CPZ-BSA, and CPZ polyclonal antibody had been produced.The CPZ polyclonal antibody the high titers of 5000 by indirect ELISA, sensitivity with 50% inhibitive concentration (IC50)of CPZ-BSA-EDC number 40 is 43.65ng/ml,mixedanhydride-CPZ-BSAis 87.65ng/ml.The CPZ polyclonal antibody with cross-reactivity to substance , promethazine,chloprothixene,perphenazine,fluphenazine hydrochloride,thioridazine hydrochloride is 26.67%,23.87%,28.8%,42.79%,31.1%.The detection limit was 4.5 ng/ml, Resuide analysis of milk, the recovery of chlorpromazine was from 65～114.6%.