Analysis of quinidine for fish tissues using single drop microextraction (SDME) coupled with atmospheric pressure matrix assisted laser desorption/ionization mass spectrometry (AP-MALDI-MS) are reported. Optimization conditions; such as extraction solvent, extraction time, pH of the aqueous solution, salt additions (NaCl), stirring rate, matrix type and concentration are investigated. Linear dynamic range (µM), limit of detection, relative recovery%, and enrichment factor are 0.08-9.2, 0.05, 94.8±3.1-98.5±3.3%, 4.34±0.28-4.40±0.30, respectively. SDME-AP-MALDI-MS shows good intraday and interday reproducibility.