Matsumoto K, Kikuchi H, Iri H, Takahasi H, Umino M
Department of Clinical Laboratories, School of Medicine, Keio University, Tokyo, Japan.
J Chromatogr. 1988 Mar 18;425(2):323-30. doi: 10.1016/0378-4347(88)80036-7.
The automated determination of theophylline and related compounds in human serum by column-switching high-performance liquid chromatography, including direct injection of serum samples, is described. TSK pre-column BSA-ODS and TSK gel ODS-80TM were used in the pre-column and analytical column, respectively. Serum samples of 20 microliters were directly injected on to the pre-column. After washing out serum proteins from the pre-column with 0.1 M NaH2PO4 at a flow-rate of 1.0 ml/min for 3.5 min, the effluent from the pre-column was introduced on to the analytical column by a column-switching device. The analysis was performed by stepwise gradient elution using 10 and 18% methanol in 0.1 M NaH2PO4. Theophylline and nine derivatives could be determined simultaneously within 40 min. The recovery of these compounds from serum was 95-103%. The linearity (1.0-50 micrograms/ml theophylline) and reproducibility (coefficient of variation less than 2.0%) were sufficient for drug monitoring at the lower and upper limits of therapeutic concentrations of theophylline.
本文描述了通过柱切换高效液相色谱法自动测定人血清中的茶碱及相关化合物，包括直接进样血清样品。预柱采用TSK预柱BSA - ODS，分析柱采用TSK凝胶ODS - 80TM。将20微升血清样品直接进样到预柱上。用0.1M NaH2PO4以1.0毫升/分钟的流速冲洗预柱3.5分钟以去除血清蛋白后，预柱流出物通过柱切换装置导入分析柱。采用在0.1M NaH2PO4中10%和18%甲醇的梯度洗脱进行分析。茶碱及其九种衍生物可在40分钟内同时测定。这些化合物从血清中的回收率为95 - 103%。线性范围（茶碱浓度为1.0 - 50微克/毫升）和重现性（变异系数小于2.0%）足以在茶碱治疗浓度的下限和上限进行药物监测。
