Carducci C, Moretti F, Birarelli M, Antonozzi I
Department of Experimental Medicine, University La Sapienza of Rome, Italy.
J Chromatogr. 1991 Aug 16;553(1-2):149-54. doi: 10.1016/s0021-9673(01)88483-1.
In order to overcome problems related to the determination of free tryptophan in biological fluids using conventional methods, we have developed an accurate and reliable procedure based on a specific pretreatment of samples followed by a very rapid and sensitive reversed-phase high-performance liquid chromatographic analysis. The pretreatment consists of adding to the samples of a very low amount of 3 M phosphate buffer to maintain pH in the physiological range followed by ultrafiltration. The precision, reproducibility and sensitivity of our method were also evaluated. The recovery of each amino acid was greater than 92%. The use of a microbore column allows the detection of up to 0.2 pmol/microliter of amino acid. The method has been applied to the analysis of samples obtained from 25 normal and 10 phenylketonuric subjects.
为了克服使用传统方法测定生物体液中游离色氨酸所涉及的问题，我们开发了一种准确可靠的方法，该方法基于对样品进行特定预处理，然后进行非常快速且灵敏的反相高效液相色谱分析。预处理包括向样品中加入极少量的3M磷酸盐缓冲液以维持pH在生理范围内，随后进行超滤。我们还评估了该方法的精密度、重现性和灵敏度。每种氨基酸的回收率均大于92%。使用微径柱能够检测至0.2 pmol/微升的氨基酸。该方法已应用于对25名正常受试者和10名苯丙酮尿症患者所采集样品的分析。
