高效液相色谱法同时测定人血清中茶碱、二羟丙茶碱、多索茶碱的浓度

目的:建立人血清中茶碱、二羟丙茶碱和多索茶碱血药浓度同时测定的高效液相色谱法并应用于患者的个体化治疗。方法:采用蛋白沉淀法对血样进行预处理,以咖啡因为内标。色谱条件:Eclipse XDB-C₁₈(4.6 mm×150 mm,5 μm)色谱柱,流动相为甲醇-水梯度洗脱,流速为1.0 ml·min^-1,检测波长273 nm,柱温为30 ℃。结果:茶碱在1.47~47.00 μg·ml^-1范围内线性良好(r=0.998),二羟丙茶碱在1.28~40.80 μg·ml^-1范围内线性良好(r=0.999),多索茶碱在1.44~46.00 μg·ml^-1范围内线性良好(r=0.999)。茶碱、二羟丙茶碱和多索茶碱的平均绝对回收率均达到95%左右,平均相对回收率分别为100.34%~105.48%,97.09%~103.13%和97.61%~101.39%,日内和日间RSD均小于10%。结论:本方法简便、快速、灵敏,可用于茶碱、二羟丙茶碱和多索茶碱的临床血药浓度监测及药动学研究。

Simultaneous determination of theophylline,diprophylline and doxofylline in serum by HPLC

OBJECTIVE To establish an HPLC method for simultaneous determination of theophylline,diprophylline and doxofylline in serum and apply it to individualized treatment.METHODS Blood samples were pretreated by using protein precipitation with caffeine as an internal standard.Analytical column was Eclipse XDB-C₁₈ (4.6 mm×150 mm,5 μm),mobile phase was methanol-water solution in gradient mode at a flow rate of 1.0 ml·min^-1.Detection wavelength was set at 273 nm and temperature was kept at 30 ℃.RESULTS Theophylline,diprophylline and doxofylline showed good linear relationships in ranges of 1.47-47.00 μg·ml^-1 (r=0.998),1.28-40.80 μg·ml^-1 (r=0.999) and 1.44-46.00 μg·ml^-1 (r=0.999),respectively.Average absolute recoveries of theophylline,diprophylline,doxofylline were above 95%,and relative recoveries were 100.34%-105.48%,97.09%-103.13% and 97.61%-101.39%,respectively.Intra-day and inter-day RSD values were less than 10%.CONCLUSION The method is simple,quick and sensitive for drug monitoring and pharmacokinetics study on theophylline,diprophylline and doxofylline.