Distribucija i određivanje niklozamida u tkivima i organima žarana (Cyprinus carpio)primenom HPLC-DAD i MS-M-a
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Niclosamide is a medication used to treat tapeworm infestation in animals and humans. Its most important use in common carp (Cyprinus carpio) is for the treatment of Bothriocephalus acheilognathi, which is a very common parasite in this fish species. The aim of this study was to optimize method determination of niclosamide (NIC) and its metabolite 2-chloro 4-nitro aniline (CNA) and 5-chloro salycilic acid (CSA) in the muscles, liver, kidneys and skin of common carp fingerlings by using HPLC-DAD-FLD-MS-MS technique. The quantification limits determined on the HPLC-DAD-FLD for the nicolosamide and the metabolites were in the range of 1,5-1,6 μg/kg for the NIC; 3,3-3,7 μg/kg for CSA and 3,3-3,6 μg/kg for CNA. LOQ, quantification limits for nicosamide and metabolites determined on LC-MS-MS were 0,4 μg/kg for NIC, while CSA ranged in the range of 0,6-1 μg/kg, and for CNA in the range from 0,5 to 0,6 μg/kg. The detection limit for NIC, CSA and CNA determined on HPLC-DAD-FLD was 0,5 μg/kg; 1 μg/kg and 1 μg/kg. The detection limits for NIC, CSA and CNA determined on LC-MS-MS were 0.1 μg / kgThe fish for the experiment were obtained from Kapetanski Rit fish pond, and were acclimated to test conditions at 20,5 ± 1°C. Common carps with an average mass of 60 ± 10 g were treated with niclosamide concentration of 2 g/kg of feed during five consecutive days. At the end of the treatment, the levels of niclosamide residues were determined using a high performance liquid chromatography (HPLC) analysis during over 13 days. The mean values of niclosamide concentrations in the muscles ranged from 27,7μg/kg starting from the first day to <0,5 μg/kg on the 11th day in muscles. The niclosamide concentration in the liver were found to be 51,5 μg/kg the first day and decreased proportionally to <0,5 μg/kg on the 13th day; in kidneys concentration were in range of 29,4 μg/kg to <0,5 μg/kg 13th day and in skin from 77,8 μg/kg starting first day to <0,5 μg/kg at 5th day. CNA level in the liver of treated common carps amounted to 170,1 μg/kg on the first day and continuously declined until the 13thday when recorded values were <1 μg/kg, in kidneys from 39,4 μg/kg to <1 μg/kg, in muscles concentration of CNA was in range 14,5 ěg/kg to <1 ěg/kg 11th day and in skin from 3,2 ěg/kg to <1 ěg / kg. CSA concentrations in the liver reached a maximum level of 11,5 ěg/kg on the 7th day and fell to <1 ěg/kg on the 13th day, and in kidneys reached maximum level of 8,4 ěg/kg on the 5th day and fell to <1 ěg/kg on the 13th day. CSA was not detected in skin. Data obtained during this study provided information about the concentration and fate of niclosamide and it's residues CNA and CSA in the liver and muscles of common carp treated orally.