Measurement of nicotine in blood of birds by reverse phase HPLC

CID, F.D.; CHEDIACK, J.G.; ENRIQUE CAVIEDES VIDAL

Mendoza

Congreso; Reunión Anual de la Sociedad de Biología de Cuyo; 2005

MEASUREMENT OF NICOTINE IN BIRD BLOOD BY REVERSE PHASE HPLC: PRELIMINARY DATA ON NICOTINE ELIMINATION IN PIGEONS Cid FD, Chediack J G, Caviedes- Vidal E. Lab Biol., Univ Nac San Luis, 5700 San Luis. E-mail fabricio.cid@gmail.com Wild animals are exposed to xenobiotics, which are absorbed in the intestine by transcellular and paracellular routes, depending on the size and charge of the molecule, its water solubility and the presence of transporters on the apical membrane of enterocytes. Nicotine is a water-soluble toxin present in several plants consumed by birds. An interesting problem in ecotoxicology is to know how much nicotine is absorbed by the bird intestine. We present here an adapted technique to assess nicotine in blood samples of pigeons (Columba livia) using RP-HPLC with electrochemical detection based on the method of Mahoney and Al-Delaimy (2001) designed to determine the toxin in hairs. As an application example, the kinetics of nicotine elimination from blood after intramuscular injection was described. Nicotine was extracted from plasma samples with dichloromethane. The solution was evaporated and the residue, in phosphate buffer, was analyzed by HPLC with a reverse phase C18 column in isocratic condition and a solvent flow of 1 mL1min (50 mrnol1L KH,PO,: 2.5% MeOH: 4% acetonitrile; pH 4.8). An electrochemical detector was used (oxidation potential: +1.10 V, sensibility: 200 nA.V-I). The detection limit of nicotine was 1.2 ng/mL of plasma. Nicotine elimination from blood shows apparent first order kinetics. This method is suitable for assaying low nicotine levels in plasma.