Determination of Doxofylline Concentration in Human Serum by Online Two-dimensional Column Switching-HPLC / 中国药房

ABSTRACT

OBJECTIVE:To establish the method for the determination of doxofylline in human serum. METHODS:Online two-dimensional column switching-HPLC was adopted to determine the concentration of doxofylline in human serum. First-dimensional chromatographic column was Waters C18column,and middle column was SC2. First-dimensional and second-dimensional column mobile phrase were methanol-water(70 30,V/V). The detection wavelength was 273 nm,and column temperature was 40 ℃. Sample volume was 10 μL. RESULTS:The linear range of doxofylline were 0.5-50.00 μg/mL(r=0.999 9), and the detection limit was 0.01 μg/mL. The quantitative limit was 0.5 μg/mL. RSDs of intra-day and inter-day were 1.51%-1.89%and 1.52%-1.92%(n=5). The accuracy were 97.91%-104.19%(n=5). Extraction recoveries rate were 91.63%-93.44%(RSD<2.00%,n=3). RSD of matrix effect were lower than or equal to 3.01%(n=6),and RSD of stability test was lower than 5.00%(n=6). After 3 patients were given intravenous injection of Doxofylline and glucose injection(0.3 g/d)up to steady state,the serum concentrations of doxofylline were 3.23,3.35,3.68 μg/mL before medication on the next day(RSD=2.28%,2.34%, 2.14%,n=5). CONCLUSIONS:The method is simple,rapid,accurate and suitable for the determination of plasma concentration of doxofylline.