Huclová Jitka, Satínský Dalibor, Maia Tiago, Karlícek Rolf, Solich Petr, Araújo Alberto Nova
Department of Analytical Chemistry, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Heyrovského 1203, 500 05 Hradec Králové, Czech Republic.
J Chromatogr A. 2005 Sep 16;1087(1-2):245-51. doi: 10.1016/j.chroma.2004.11.055.
Restricted access material (RAM) column containing 25 microm C18 alkyl-diol support was integrated into the sequential injection analysis (SIA) manifold and the SIA-RAM system was tested for direct determination of furosemide in serum. LiChrospher ADS column based on restricted access material is proposed to direct injection of biofluids. The integration of RAM material into SIA enabled creation of a comprehensive on-line sample clean-up technique combined with fluorescence quantitation of analyte. Centrifuged and diluted serum sample was aspirated into the system and loaded onto the column using acetonitrile-water (2:98), pH 2.7. The analyte was retained on the column while proteins contained in the sample were removed to the waste without precipitation and clogging the column. Interfering substances complicating the detection were washed out by acetonitrile-water (15:85), pH 2.7 in the next step. The extracted analyte was eluted by means of acetonitrile-water (25:75), pH 2.3 to the fluorescence detector (emission filter 385 nm). The whole procedure comprising sample pre-treatment, analyte detection and column reconditioning took 20 min. The recoveries of furosemide from serum lay between 101.4 and 103.4% for three concentrations of analyte.
将含有25微米C18烷基二醇载体的受限进样材料（RAM）柱集成到顺序注射分析（SIA）流路中，并对SIA-RAM系统进行了血清中呋塞米直接测定的测试。提出了基于受限进样材料的LiChrospher ADS柱用于生物流体的直接进样。将RAM材料集成到SIA中，能够创建一种综合的在线样品净化技术，并结合分析物的荧光定量分析。将离心并稀释后的血清样品吸入系统，使用乙腈-水（2:98，pH 2.7）加载到柱上。分析物保留在柱上，而样品中含有的蛋白质则被排至废液，不会发生沉淀和堵塞柱子的情况。下一步用乙腈-水（15:85，pH 2.7）冲洗掉使检测复杂化的干扰物质。用乙腈-水（25:75，pH 2.3）将萃取的分析物洗脱至荧光检测器（发射滤光片385纳米）。整个过程包括样品预处理、分析物检测和柱再生，耗时20分钟。对于三种分析物浓度，血清中呋塞米的回收率在101.4%至103.4%之间。
