Quantitative HPLC analysis of acidic opines by phenylthiocarbamyl derivatization

A new method for the quantitative analysis of acidic opines--alanopine, strombine, tauropine, and beta-alanopine--is presented. The method is based on formation of phenylthiocarbamyl (PTC) derivatives of the acidic opines after partial purification by ion-exchange chromatography. The PTC acidic opines are separated by reverse-phase high-performance liquid chromatography (HPLC) and detected within 20 min by ultraviolet absorbance. This HPLC method gives higher sensitivity, 10-30 pmol minimum detection, and better reproducibility than the high-voltage paper electrophoresis method. There is also good agreement for the three acidic opines (alanopine, strombine, and tauropine) when compared by HPLC and electrophoresis methods. Accumulation of beta-alanopine was observed for the first time in the adductor muscle of blood shell, Scapharca broughtonii, during aerial exposure by application of the HPLC detection method.