There is a growing body of evidence that toxic organotins are making their way into terrestrial and aquatic mammals including humans. In the United States, one possible route of environmental exposure to organotins (specifically dibutyltin and triphenyltin) is via fresh surface waters and fish taken from those waters. A unique methodology was used for quantitative and speciation of the organotins. This green-chemistry method combines two extraction techniques (solid-phase extraction for waters; hexane/tropolone extraction for fish) with micro-liquid chromatography-electrospray/ion trap mass spectrometry (micro-LC-ES/ITMS) as the detection method. A small survey looking for organotins in fresh surface waters across the United States, and fish from those waters, was conducted. Various concentrations of dibutyltin and triphenyltin were detected in fresh water, ranging from nondetect to 2 ppb, and nondetect to 6 ppb, respectively. In fish dibutyltin and triphenyltin were detected from nondetect to 200 ppb, and nondetect to 400 ppb, respectively.