Simultaneous determination of plasma phenytoin and its primary hydroxylated metabolites in carbon tetrachloride-intoxicated rats by high-performance liquid chromatography.
We have developed a simple and sensitive method for the simultaneous determination of phenytoin (PHT), 5-(p-hydroxyphenyl)-5-phenylhydantoin (p-HPPH) and 5-(m-hydroxyphenyl)-5-phenylhydantoin (m-HPPH) in rat plasma by high-performance liquid chromatography. The three substances were separated on a reversed-phase column (5 microns TSK gel ODS-80TM, 250 mm x 4.6 mm I.D.) using acetonitrile-0.008 M NaH2PO4 (pH 6) (35:65, v/v) as a mobile phase at a flow-rate of 0.8 ml/min. Absorbance was monitored at 215 nm. The quantification limit was 50 ng/ml for each of PHT, m-HPPH and p-HPPH. The mean recoveries for DPH, m-HPPH and p-HPPH from plasma were 95.6 +/- 3.6, 94.5 +/- 4.2 and 98.6 +/- 2.9%, respectively.
Tanaka
E
E
Institute of Community Medicine, University of Tsukuba, Ibaraki-ken, Japan.
Sakamoto
N
N
Inubushi
M
M
Misawa
S
S
D000818:Animals; D000927:Anticonvulsants / Q000097:blood*; D002251:Carbon Tetrachloride / Q000008:administration & dosage / Q000633:toxicity*; D002851:Chromatography, High Pressure Liquid; D008297:Male; D010672:Phenytoin / Q000031:analogs & derivatives* / Q000097:blood* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013053:Spectrophotometry
D000927:Anticonvulsants; C001419:hydroxyphenytoin; C027364:5-(3-hydroxyphenyl)-5-phenylhydantoin; D010672:Phenytoin; D002251:Carbon Tetrachloride