摘要
本文从经济、实用的角度出发,建立并比较了在反相(A法)和正相(B法)色谱系统下测定血清中茶碱浓度的两种高效液相色谱法(HPLC)。A法采用了ZORBAXODS色谱柱,甲醇、醋酸盐缓冲液为流动相;B法采用ZORBAXSIL色谱柱,正已烷、氯仿、甲醇三元混合溶剂为流动相,均以β-羟乙基茶碱为内标、275nm为检测波长。测定结果表明,两法线性范围均为2.5～40.Oμg/ml,最低检测浓度为2.0μg/ml,并且均有良好的相关性(r=0.999)。目内、目间变异小于6.36％,A法和B法的回收率分别为100.1±3.68％和99.1±4.33％,分析时间分别为11min和7min,血清内源物和部分临床常用药物不干扰两法测定。两法的测定结果相关性良好,回归方程为 =0.9681×+0.1504(r=0.999,n=5)。经临床的茶碱样品测定表明,两法简便灵敏,快速实用,能满足临床日常监测血药浓度水平的需要。
Two rapid, practical methods utilizing Reversed-Plase(method A) and Nor-mal-Phase High Performance Liquid Chromatography(method B) for the quantitation of theophylline in serum are described. The same extraction procedures of sample are used for the two methods. The wavelength of UV detcctor is set at 275 nm and beta-hydroxyethyltheophylline is used as the internal standard. The linear rangs of the two methods are all from 2.5～40.0ug/ml with good correlation coefficients(0.999), the sensitivity limits are all 2.0ug/ml. The variation coefficients of within-day and day-to-day at different concentration levels are all less than 6.36%. Recoveries arc 100.4⊥ 3.68%(mcthod A) and 99.1±4.33%(method B). Chromatogram run times are ll minutes and 7 minutes. The two methods arc interference free from the endogcncous serum components and other drugs commonly coadministered with theophyllinc. There is a better correlation between the two methods with a correlation coefficient 0.999 and linear curve Y=0.9681X+0.504(n= 5) at four concentration levels. These methds have been applied o clinical therapeutic drug monitoring(TDM) in theophylline and harvest a good result.
