Determination of antihypertensive small peptides, Val-Tyr and Ile-Val-Tyr, by fluorometric high-performance liquid chromatography combined with a double heart-cut column-switching technique.

A double column-switching HPLC method with naphthalene-2,3-dialdehyde (NDA) was applied for determination of two plasma antihypertensive peptides, Val-Tyr (VY) and Ile-Val-Tyr (IVY). After a first separation on a Phe-ODS column, double heart-cuts of the retention time corresponding to NDA-VY and NDA-IVY elutions were successfully separated on an analytical ODS column: 60% acetonitrile in 0.1% trifluoroacetic acid containing 5 mM sodium 1-octanesulfonate at 1.0 mL/min. Within-run coefficients of variation were 1.73% and 4.73% for VY and IVY, respectively.