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Abstract
A sensitive and selective analysis for the determination of β-endorphin related peptides in human plasma is described. Laser induced fluorescence (LIF) reaction detection has been used for ultra trace level detection. Two different derivatization strategies, each demanding its specific sample pretreatment procedure, are described. In the first method the fluorophoric agent fluorescein-5-isothiocyanate (FITC) has been applied as pre-column derivatization reagent. In the second approach the fluorogenic agent o-phthaldialdehyde (OPA) is used in the post-column derivatization mode. The derivatives are detected by means of a home-made LIF detection system applying either the UV multiline or the intense 488 nm emission line of a water-cooled argon-ion laser. Determination limits achieved with the OPA post-column reaction are about 3 orders of magnitude lower than those obtained with FITC pre-column derivatization. With OPA post-column reaction detection and LIF detection, a determination limit of 0.4 ng/ml has been realized for the model compound des-enkephalin-γ-endorphin (β-endorphin 6–17).
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