Laser Fluorescence Detection in Microcolumn Liquid Chromatography: Application to Derivatized Carboxylic Acids

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Abstract:

A laser-induced fluorescence (LIF) detector is evaluated for use with high-efficiency liquid chromatography columns of capillary dimensions. This detector employs a cw helium-cadmium laser (325 nm, 5-10 mW) as the excitation source, together with a fused-silica capillary flowcell and filter/photomultiplier optical system. The flowcell, which is an integral part of the microcolumn, contributes from 0.06 nL2 to 0.06 μL2 variance to the chromatographic system for capillaries of 35 to 330 μm diameter, respectively. The LIF detector is capable of sensing femtogram amounts of coumarin 440, and the response is linearly related to concentration over at least eight orders of magnitude. The potential of this combination of analytical techniques is demonstrated through the analysis of multicomponent mixtures of derivatized fatty acids and prostaglandins at the pico- and subpicomole level.

Keywords: Fatty acid and prostaglandin analysis; Laser fluorescence detection; Microcolumn liquid chromatography

Document Type: Research Article

DOI: http://dx.doi.org/10.1366/0003702854249989

Affiliations: 1: Department of Chemistry, Stanford University, Stanford, California 94305; current address: Department of Chemistry, Michigan State University, East Lansing, MI 48824 2: Department of Chemistry, Stanford University, Stanford, California 94305

Publication date: September 1, 1985

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