Determination Of Nicotine And Its Metabolites Accumulated In Fish Tissue Using Quechers And Hydrophilic Interaction Liquid Chromatography And Tandem Mass Spectrometry
The quantitative determination of nicotine and its major metabolites (cotinine and anabasine) in fish tissue was performed using hydrophilic interaction liquid chromatography (HILIC) coupled with tandem mass spectrometry. Marine and freshwater fish were purchased from local grocery stores and were prepared based on the well-known QuEChERS (quick, easy, cheap, effective, rugged, safe) sample preparation protocol. There were significant suppressions on measured nicotine signals (10%) due to the matrix effects from marine fish, but no obvious effects on freshwater fish signals. Method validation was incorporated with internal standards and carried out with matrix-matched calibration. The detection limits for nicotine, cotinine, and anabasine were 9.4, 3.0, and 1.5 ng/g in fish, respectively. Precision was acceptable and less than 9% RSD at low, mid, and high concentrations. Acceptable and reproducible extraction recoveries (70 - 120%) of all three compounds were achieved, except for anabasine at low concentration (61%). This developed method offers a fast, easy, and sensitive way to evaluate nicotine and its metabolite in fish tissues.