Determination of N- (trans-4-isopropylcyclohexylcarbonyl) -D-phenylalanine in human plasma by solid-phase extraction and column-switching high-performance liquid chromatography with ultraviolet detection

Ono, I.; Matsuda, K.; Kanno, S.

Journal of Chromatography. B, Biomedical Applications 678(2): 384-387


ISSN/ISBN: 1572-6495
PMID: 8738049
DOI: 10.1016/0378-4347(95)00511-0
Accession: 008443919

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A column-switching high-performance liquid chromatography method with ultraviolet detection at 210 nm has been developed for the determination of N-(trans-4-isopropylcyclohexylcarbonyl)-D-phenylalanine (AY4166, I) in human plasma. Plasma samples were prepared by solid-phase extraction with Sep-Pak Light tC18, followed by HPLC. The calibration graph for I was linear in the range 0.1-20 micrograms/ml. The limit of quantitation of I, in plasma, was 0.05 microgram/ml. The recovery of spiked I (0.5 microgram/ml) to drug-free plasma was over 92% and the relative standard deviation of spiked I (0.5 microgram/ml) compared to drug-free plasma was 4.3% (n = 8).