High-performance liquid chromatographic method for quantitating plasma levels of amiloride and its analogues

• Published in: Journal of chromatography. Biomedical applications 1992-11, Vol.582 (1), p.217-223
• Main Authors: Alliegro, Mary Anne, Dyer, Kimberly D., Cragoe, Edward J., Glaser, Bert M., Alliegro, Mark C.
• Format: Article
• Language: English
• Subjects: Analysis of complex biological substances; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Blood and biological fluids; Fundamental and applied biological sciences. Psychology
• ISSN: 0378-4347
• DOI: 10.1016/0378-4347(92)80322-H

An assay for amiloride was devised for efficient use with the wide variety of analogues available. Amiloride was extracted from 1-ml plasma samples by elution from a C 8 preparative column with 6% acetonitrile—45% methanol—5.4% acetic acid, adjusted to pH 4.0 with trimethylamine. Samples were lyophilized, resuspended in 50% methanol, filtered through 0.22-μm Spin-X cartridges, applied to a reversed-phase C 18 column, and eluted in a 0–50% acetonitrile gradient in 0.4% acetic acid, pH 4.5 (1.2 ml/min). Detection by ultraviolet absorbance at 360 nm was linear from 1 to 1000 ng. Versatility of the method was demonstrated with the analogues benzamil, 6-hydro-, 6-iodo-, 5-hexamethylene-, and 5-chlorobenzyl-2',4'-dimethylbenzylamiloride.