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

D. Roy, W. R. Snodgrass

Research output: Contribution to journalArticle

7 Citations (Scopus)

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 beta-glucuronidase, 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 isopropranol:water (20:80) at a flow rate of 1.4 ml/min and monitored at 235 mm. 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.

Original languageEnglish (US)
Pages (from-to)57-62
Number of pages6
JournalTherapeutic Drug Monitoring
Volume11
Issue number1
StatePublished - 1989

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High performance liquid chromatography
Phenytoin
Metabolites
Microsomes
Animals
High Pressure Liquid Chromatography
Urine
Chromatographic analysis
Glucuronidase
Biological materials
Chromatography
Durability
Flow rate
Water
Research
hydroxyphenytoin
ethyl acetate
octadecyl silane-bonded silica
catechol
phenytoin dihydrodiol

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Health, Toxicology and Mutagenesis
  • Pharmacology (medical)
  • Public Health, Environmental and Occupational Health
  • Pharmacology
  • Toxicology

Cite this

Isocratic separation of phenytoin metabolites by high-performance liquid chromatography from human and animal microsomes and urine. / Roy, D.; Snodgrass, W. R.

In: Therapeutic Drug Monitoring, Vol. 11, No. 1, 1989, p. 57-62.

Research output: Contribution to journalArticle

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