Huber J F, Lamprecht G
Institute for Analytical Chemistry, University of Vienna, Austria.
J Chromatogr B Biomed Appl. 1995 Apr 21;666(2):223-32. doi: 10.1016/0378-4347(94)00568-p.
An automated two-dimensional HPLC method for the determination of neopterin in serum is described. Neopterin is separated from proteins on a short octadecylsilica column by size exclusion and from the majority of the other serum components by adsorption. The fraction containing neopterin is transferred by column switching to a solvent-generated cation-exchange column using dodecylsulfonic acid as surface activator. Parameters influencing the separation performance and sensitivity of the fluorescence detection are discussed. The efficiency of the cleaning of the first column was optimized. The method was validated. It achieves a precision of 1% (R.S.D.) and a detection limit of about 0.3 nmol/l. The accuracy is nearly 100%. The method allows a high sample throughput, requiring 15 min per sample.
本文描述了一种用于测定血清中蝶呤的自动化二维高效液相色谱法。蝶呤通过尺寸排阻在短十八烷基硅胶柱上与蛋白质分离，并通过吸附与大多数其他血清成分分离。含有蝶呤的馏分通过柱切换转移到以十二烷基磺酸为表面活性剂的溶剂生成阳离子交换柱上。讨论了影响荧光检测分离性能和灵敏度的参数。优化了第一根柱的清洗效率。该方法经过验证。其精密度为1%（相对标准偏差），检测限约为0.3 nmol/l。准确度接近100%。该方法允许高通量进样，每个样品需要15分钟。
