A new enzyme immunoassay for aconitine and its application to quantitative determination of aconitine levels in plasma.

摘要: A reliable enzyme immunoassay (EIA) method was developed for quantitative determination of aconitine with high sensitivity and specificity. The bovine serum albumin (BSA)- β-galactosidase (β-Gal) conjugates as immunogens enzyme-labeled antigens were prepared by coupling their proteins succinic acid (short chain length; n=2, where n represents the number methylene units) hexadecanedioic (long n=14) hemiesters benzoylaconine through respective N-hydroxysuccinimide esters intermediates. Two types BSA-conjugates short long chains repeatedly injected into rabbits to obtain anti-aconitine antisera (As1 As2, respectively). All combinations β-Gal-labeled LAg1 (n=2) LAg2 (n=14) As1 As2 showed in a range 0.1—1.0 ng. Although combination antiserum specificity aconitine, highly specific both mesaconitine. When intravenously administered rats, concentration plasma remarkably decreased within first 60 min, then gradually declined, suggesting two-compartment pharmacokinetic model (Vc 0.41±0.09 l/kg, Vdss 1.7±0.4 CLtot 10±2 ml/min · kg, AUC0—4800 2055±294.3 ng · min/ml). Following oral administration rats at two doses 0.1 1.0 mg/kg b.w., maximum concentrations (Cmax) 0.73±0.08 3.3±0.6 ng/ml times 45±9 150±52 respectively, AUC0—1440 values 130±4 1600±270 ng · min/ml. bioavailability (F) determined be 0.013, only 1.3% orally absorbed body fluid.