HPLCFLD

HPLCFLD stands for high-performance liquid chromatography with fluorescence detection. It is an analytical technique that combines the separation capability of HPLC with the sensitivity of fluorescence detection, using detectors that measure emitted light from fluorescent analytes or from derivatized compounds.

In operation, a sample is injected into an HPLC system and separated on a chromatographic column by a mobile phase. As analytes elute, they pass through a fluorescence detector, which excites them with light at specified wavelengths and records the emitted fluorescence. Only compounds that are intrinsically fluorescent or have been derivatized to fluoresce are detected, allowing very low limits of detection in many cases.

A fluorescence detector typically comprises an excitation light source (such as a xenon lamp or LEDs), an emission monochromator or optical filters, and a photodetector. Correct selection of excitation and emission wavelengths is essential for optimizing sensitivity and selectivity for the target analyte.

Derivatization is common when target compounds lack native fluorescence. Reagents such as o-phthalaldehyde, fluorescamine, or other fluorescent tags can convert amino acids, amines, or other species into fluorescent derivatives, enabling their detection by HPLCFLD.

Applications of HPLCFLD include clinical chemistry (e.g., amino acids and neurotransmitters), pharmaceutical analysis, environmental monitoring (pesticides and certain contaminants), and food safety (flavoured or fluorescent additives). It offers high sensitivity, wide dynamic range, and low background compared with UV detection, but is limited to fluorescent analytes and can require additional sample preparation and method development.