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Single drop microextraction has become a widespread liquid/liquid microextraction technique owing to its simplicity, high preconcentration factor and low consumption of organic solvents in the extraction due to direct introduction of the very low volume of extract into the analytical system. Crucial features ensuring excellent repeatability of single drop microextraction include: solvent volume, solvent type, sample agitation, salts addition, and pH. The influence of sample quality on the microdrop volume and agitation type was studied. Effect of the sample matrix, such as water, acid content samples (orange juice), sample containing alcohol (plum brandy) and protein content sample (milk), on the microdrop hold-up was also investigated. For water analysis, several organic solvents such as chloroform, dichloromethane, tetrachloromethane, tetrachloroethane and chlorobenzene were tested; last three mentioned were suitable for SDME experiments. For milk samples analysis, chlorobenzene microdrop was found to be optimal; advantage of salt addition has been reported. For orange samples, 1:10 dilution was suggested with stable microdrop volumes (toluene) of up to 6 µL for lower stirring rates (100 rpm and 250 rpm). For alcohol-content samples, the change of alcohol percentage of real-life samples had to be considered. A strong influence of the matrix quality on the microdrop stability has been proven.