Simultaneous determination of anticonvulsants in serum by HPLC using dual column-dual detection

• Published in: BUNSEKI KAGAKU Vol. 38; no. 11; pp. T170 - T173
• Main Authors: FUJITA, Tomio, GAMOH, Keiji, MURAKITA, Hiroyuki
• Format: Journal Article
• Language: English; Japanese
• Published: Tokyo The Japan Society for Analytical Chemistry 1989; Japan Science and Technology Agency
• Subjects: determination of anticonvulsants; dual column-dual detection technique; reversed phase HPLC
• ISSN: 0525-1931
• DOI: 10.2116/bunsekikagaku.38.11_T170

We present a rapid and conventional HPLC method for the simultaneous determination of anticonvulsants, ESM, PM, PB, CBZ, PHT, NZP, CZP and DZP in serum using the dual column-dual detection technique. The system consists of two flow lines connected through the flow changeover valve, and is controlled by a time program using a microprocessor. These compounds were separated by the reversed phase mode with isocratic elution. The analytical procedure was as follows; a serum extract was injected into the first analytical column. After 0.5 min, the flow line was changed to the dotted line by turning the changeover valve. In this stage, weakly retained compounds, such as ESM, PM and PB were eluted into the second column, and monitored at 220 nm. After 7 min, the flow line was returned to the original line. CBZ, PHT and benzodiazepines were separated in the column 1 and monitored at 254 nm. Both detectors 1 and 2 are connected to data processor and thus analytical results can be calculated at the same time. A single analysis was completed in about 20 min. The calibration curves for ESM and NZP were linear over the ranges 5.768.4 μg/ml and 17160 ng/ml, respectivery. Recovery of the drugs from spiked serum were from 98.9±0.9% (ESM) to 78.6±4.9 (NZP). The present method was used for the determination of anticonvulsants in the serum of an epileptic patient.