Journal
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
Volume 38, Issue 5, Pages 910-917Publisher
ELSEVIER
DOI: 10.1016/j.jpba.2005.01.049
Keywords
miniaturization; capillary HPLC-NMR; hyphenated techniques; carotenoids
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The development of miniaturized hyphenated systems such as capillary high-performance liquid chromatography-and nuclear magnetic resonance spectroscopy (HPLC-NMR) remains challenging in the field of structure elucidation. In combination with a highly specific sample preparation technique, matrix solid-phase dispersion (NISPID), and a highly selective C-30 reverse phase HPLC-NMR enables the identification of small amounts of natural compounds. Here, the investigation of five carotenoids in a standard solution and two carotenoids from a spinach sample demonstrate the potential of this new development. The separation of the carotenoids is performed with self-packed fused-silica capillaries with a binary solvent gradient consisting of acetone and water. The miniaturized system allows the use of fully deuterated solvents for on-line HPLC-NMR coupling. The H-1 NMR spectra of the various carotenoids obtained in stopped-flow mode gave a high signal-to-noise ratio with a sample amount in the low nanogram range. All necessary parameters for structure elucidation such as multiplet structure, coupling constants and integration values can be detected unambiguously. (c) 2005 Elsevier B.V. All rights reserved.
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