摘要
采用高效液相色谱 紫外检测法分离测定了血清中的芳香族氨基酸。采用的色谱柱为WatersNova PakC18柱(4μm,150mm×3 9mmi d ),流动相为乙腈 水(体积比为6∶94,pH3 4)溶液,流速为1 0mL/min,检测波长为215nm。血清标本经5%(体积分数)高氯酸溶液去除蛋白质后取上清液直接进样,10min内完成测定。探讨了流动相的pH及其有机相的比例、蛋白质沉淀剂以及检测波长等因素对分离度和灵敏度的影响。考察了其他10余种氨基酸、多巴胺类等物质对目标组分检测的干扰。该方法的日内、日间测定的相对标准偏差均小于5%,芳香族氨基酸测定的线性范围、检测限、回收率等均满足临床检验的要求。结果表明:该方法简便、快速、准确、可靠,适用于临床和科研工作。
A high performance liquid chromatographic method, coupled with ultraviolet detection (HPLCUV), was developed for the determination of aromatic amino acids in serum. It employed a Waters NovaPak C18 column (4 μm, 39 mm id×150 mm) and a mobile phase of acetonitrilewater (6∶94, v/v; pH 34) at a flow rate of 10 mL/min. Serum samples were first precipitated with a 5% perchloric acid solution, then centrifuged to remove protein residuals and finally assayed by HPLC with UV detection at 215 nm for 10 min. The effects of pH and the organic composition of the mobile phase, the protein precipitants and the wavelength of detection on method sensitivity and separation were studied. Interferences of other amino acids and dopamines were also investigated. Intraday and interday variations of the method were all within 5%. All acceptance criteria for linearity range, limit of detection and recovery were met in compliance with established clinical study requirements. It has been demonstrated that the method is simple, fast, accurate, and convenient, and suitable for routine.
出处
《色谱》
CAS
CSCD
北大核心
2003年第2期154-157,共4页
Chinese Journal of Chromatography
