在线二维柱切换-高效液相色谱法测定人血清中多索茶碱浓度

Determination of Doxofylline Concentration in Human Serum by Online Two-dimensional Column Switching-HPLC

目的:建立测定人血清中多索茶碱浓度的方法。方法:采用在线二维柱切换-高效液相色谱法测定人血清中多索茶碱浓度,一维色谱柱为Waters C_(18),中间柱为SC2,一维、二维柱流动相均为甲醇-水(70∶30,V/V),检测波长为273 nm,柱温为40℃,进样量为10μL。结果:多索茶碱在0.5~50.00μg/m L质量浓度范围内与峰面积线性关系良好(r=0.999 9),检测限为0.01μg/m L,定量限为0.5μg/m L,日内、日间精密度的RSD分别为1.51%~1.89%和1.52%~1.92%(n=5),准确度为97.91%~104.19%(n=5),提取回收率为91.63%~93.44%(RSD<2.00%,n=3),基质效应RSD≤3.01%(n=6),稳定性的RSD<5.00%(n=6)。3例患者静脉输注多索茶碱葡萄糖注射液(0.3 g/d)达稳态后,次日给药前静脉血清中多索茶碱的浓度为3.23、3.35、3.68μg/m L,RSD为2.28%、2.34%、2.14%(n=5)。结论:该方法简便、快速、准确,适用于临床多索茶碱的血药浓度检测。

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. Firstdimensional chromatographic column was Waters C_（18） column, and middle column was SC2. First-dimensional and seconddimensional 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/m L（r=0.999 9）,and the detection limit was 0.01 μg/m L. The quantitative limit was 0.5 μg/m L. 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%（RSD2.00%,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/m L 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.