Development of an enzyme-linked immunosorbent assay for the quantification of amiodarone.
A sensitive and specific enzyme-linked immunosorbent assay for an antiarrhythmic drug, amiodarone (AMI), was developed, which is capable of measuring levels as low as 16 ng/ml. Anti-AMI antibody was obtained by immunizing rabbits with an antigen conjugated with bovine serum albumin using diazotized 4-amino-1-(2-diethylaminoethoxy)-2,6-diiodobenzene. Enzyme labeling of AMI with beta-D-galactosidase was similarly performed using a diazotized 4-amino-1-(2-diethylaminoethoxy)-2,6-diiodobenzene. This enzyme-linked immunosorbent assay was specific for AMI and showed a very slight cross-reactivity (1.25%) with its major metabolite, mono-N-desethylamiodarone. The values of the AMI concentrations measured by this assay were in good correlation to those by HPLC. Its analytical applicability was demonstrated by a kinetic study with human liver microsomes. The enzyme-linked immunosorbent assay should be a valuable tool in therapeutic drug monitoring and pharmacokinetic studies of AMI.
Saita
Tetsuya
T
Faculty of Hospital Pharmacy, Saga Medical School, Japan. saita@post.saga-med.ac.jp
Fujito
Hiroshi
H
Mori
Masato
M
D000638:Amiodarone / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005069:Evaluation Studies as Topic; D005260:Female; D006801:Humans; D008862:Microsomes, Liver; D011817:Rabbits
D000638:Amiodarone