S-Nitroso compounds have received much attention in biological research. In addition to their role as nitric oxide donors, there is growing evidence that these compounds are involved in signaling processes in biological systems. Determination of S-nitrosylated proteins is of great importance for fundamental biological research and medical applications. The most common method to assay biological S-nitroso compounds is to chemically or photochemically reduce SNO functional groups to release nitric oxide, which is then entrained in an inert gas stream and detected, usually through chemiluminescence. We report a method of S-nitroso compound detection using cavity ring-down measurements of gaseous NO absorbance at 5.2 μm. The proposed method, in contrast to the chemiluminescence-based approach, can be used to distinguish isotopic forms of NO. We demonstrated sensitivity down to ∼2 pmol of S(14)NO groups and ∼5 pmol of S(15)NO groups for S-nitroso compounds in aqueous solutions. The wide dynamic range of cavity ring-down detection allows the measurement of S-nitroso compound levels from pico- to nanomole amounts.