Experimental methodology of simultaneous determination of carbamazepine, phenytoin, and phenobarbital in serum by high-performance liquid chromatography / 中国医师杂志

ABSTRACT

Objective To establish a high-performance liquid chromatography (HPLC) method with diode array detection to simultaneously determine carbamazepine,phenytoin,and phenobarbital in serum.Methods Extraction solvent (800μl ethylene acetate) and sample (0.2 ml) was mixed,extracted for 2 min,and centrifuged (3500 r/min,4 minutes).A volume (600 μl) of extract liquor was volatilized to dryness in water bath with the volatilization temperature 75 ℃,then was redissolved with 1.0 ml mobile phase.Analysis conditions was column temperature 30°,mobile phase (methanol∶ water =40∶60),and detection wavelength of 254 nm.Three metabolites were effectively separated.Results Under the optimized condition,calibration curves of three metabolites were linear in the ranges of (1.52 ～ 120 mg/L) and the correlation coefficients were not less than 0.999.The detection limits (S/N =3) were in the range of 0.4 ～ 1.5 mg/L.The spiked recoveries were in the range of 91.3％ ～ 111％ with relative standard deviations (RSD) less than 5％.Conclusions The optimal pretreatment condition for the sample was established.The chromatographic separation and the detection condition were optimized.The method was sensitive and accurate,and could meet the need of monitoring serum drug concentration.