Isocratic separation of phenytoin metabolites by high-performance liquid chromatography from human and animal microsomes and urine Article

Roy, D, Snodgrass, WR. (1989). Isocratic separation of phenytoin metabolites by high-performance liquid chromatography from human and animal microsomes and urine . 11(1), 57-62. 10.1097/00007691-198901000-00012

cited authors

  • Roy, D; Snodgrass, WR

authors

abstract

  • A rapid and simple high-performance liquid chromatography analytical method is described for the quantitative determination of phenytoin and five of its metabolites—phenytoin dihydrodiol, catechol, methoxycatechol, para-hydroxyphenytoin, and meta-hydroxyphenytoin—in biological materials. Following ethyl acetate extraction and incubation with betaglucuronidase, samples were passed through a C18 Sep-Pak cartridge. The eluate was chromatographed on a reverse-phase 10 cm C18 Radial Nova-Pak (5 µm) column using a mobile phase of isopropanol: Water (20:80) at a flow rate of 1.4 ml/min and monitored at 235 nm. Total chromatographic analysis time was 23 min, with complete baseline resolution of all metabolites. Five different columns and 10 different mobile phase conditions were studied to determine the best system. The 10-cm C18 Radial Nova-Pak (5 µm) gave the best resolution, reproducibility, and durability. This method should prove applicable for clinical use as well as for research purposes. © 1989 Raven Press, Ltd.

publication date

  • January 1, 1989

Digital Object Identifier (DOI)

start page

  • 57

end page

  • 62

volume

  • 11

issue

  • 1