224 nm Deep-UV laser for native fluorescence, a new opportunity for biomolecules detection.

A new highly sensitive and compact 224 nm laser-induced native fluorescence (LINF) detector was developed using a new generation of deep-UV laser and an innovating elliptical flow cell. The use of deep-UV excitation at 224 nm allows to achieve fluorescence detection of an important range of molecules containing a single aromatic ring. The LINF detector was first evaluated in liquid chromatography. An improvement of a factor 500 over a conventional fluorimeter is reached with a limit of detection (LOD) of 1.5 pmole for ibuprofen. LODs were in the nanomole range for phenylalanine and in the picomole range for tyrosine and tryptophan. The LINF detector is able to detect the same levels of peptides concentrations as an ESI-ion trap spectrometer used in scan mode. In this application, LINF outperforms the UV detection at 214 or 254 nm and could be used with different additives with no noticeable effect on the detection.