Analytical Sciences
Online ISSN : 1348-2246
Print ISSN : 0910-6340
ISSN-L : 0910-6340
Determination of Antihypertensive Agent Cromakalim in Human Plasma by High Performance Liquid Chromatography Employing a Column-Switching Technique
Shinobu KUDOHHiroshi NAKAMURA
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1989 Volume 5 Issue 1 Pages 39-42

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Abstract

A high performance liquid chromatographic (HPLC) method employing a column-switching technique was developed to determine the plasma levels of Cromakalim, a novel antihypertensive agent. The plasma sample is first introduced onto a precolumn of two-phase gel where the hydrophobic drug, having a chroman skeleton, is well retained while biological macromolecules such as proteins and nucleic acids are not. The retained Cromakalim is subsequently transferred to an analytical column of octadecyl silica (ODS) using a column-switching technique, followed by fluorescence detection (Ex 254nm, Em 306nm). This system has realized a highly sensitive determination of Cromakalim in human plasma without prolonged pretreatment procedures such as solvent extraction. In tests, calibration curves were linear over the range 0.3 to 20ng ml-1 with a detection limit of 0.07ng ml-1. The relative standard deviations (n=3 each) for spiked plasma samples at 1 and 20ng ml-1 were 3.33 and 1.21 %, respectively.

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© The Japan Society for Analytical Chemistry
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