It is undeniable that hydrogen bonding plays a fundamental role in all biological systems. Not only is hydrogen bonding between water and bio-molecules important, but the more complex supramolecular structures of DNA, proteins and enzymes are formed, in part, by virtue of hydrogen bonding. The short-strong hydrogen bond (SSHB) is a critical component of a variety biological systems, where a SSHB is formed between a donor and acceptor, like all hydrogen bonds, with the donor-acceptor distance typically on the shorter end of the hydrogen bonding range. Although the SSHBs between the molecules are often located by NMR and probed in the solid state by diffraction techniques, it is exceedingly rare that these bonds are probed by direct structural techniques in non-aqueous solution, the mileu in which many SSHBs are most important.