Determination of concentrations of flecainide in human serum by high-performance liquid chromatography on a fluorocarbon-bonded silica gel column.
An optimized method for the determination of flecainide in serum is presented. Extraction using a solid-phase C18 column and chromatography on a stabilized fluorocarbon-bonded silica gel column effectively separate flecainide from an internal standard (a positional isomer of flecainide). The HPLC apparatus and conditions were as follows: analytical column, Fluofix 120N; sample solvent, 20 microl; column temperature, 40 degrees C; detector, Shimadzu RF-5000 fluorescence spectrophotometer (excitation wavelength = 300 nm, emission wavelength = 370 nm); mobile phase, 0.06% phosphoric acid containing 0.1% tetra-n-butyl ammonium bromide-acetonitrile (75:25, v/v); flow-rate, 1.0 ml/min. The standard curves for flecainide were linear in the concentration range examined (10-2000 ng/ml). The regression equation was y = 0.08+0.0078x (r = 0.9998). The minimum detectable amount of flecainide was approximately 5 ng/ml. In the within-day study, the precision coefficients of variation were 2.66, 2.18, 2.54, 2.72, 2.88, 2.24, and 3.29% for the 10, 50, 100, 200, 500, 1000, and 1500 ng/ml standards, respectively. The absolute recovery rates of flecainide at each concentrations were 94-100%. The method described provides analytical sensitivity, specificity and reproducibility suitable for both biomedical research and therapeutic drug monitoring.
Obayashi
M
M
Department of Pharmacy, Omori Hospital, Toho University School of Medicine, Tokyo, Japan.
Kosugi
T
T
Yamazaki
J
J
Matsumoto
Y
Y
Fukuoka
M
M
Matsumoto
M
M
D000889:Anti-Arrhythmia Agents / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005424:Flecainide / Q000097:blood*; D005466:Fluorocarbons / Q000737:chemistry; D006801:Humans; D015203:Reproducibility of Results; D058428:Silica Gel; D012822:Silicon Dioxide / Q000737:chemistry; D013050:Spectrometry, Fluorescence
D000889:Anti-Arrhythmia Agents; D005466:Fluorocarbons; D058428:Silica Gel; D012822:Silicon Dioxide; D005424:Flecainide