Satínský Dalibor, Serralheiro Hugo S, Solich Petr, Araújo Alberto N, Montenegro Maria C B S M
Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Heyrovského 1203, Hradec Králové 500 05, Czech Republic.
Anal Chim Acta. 2007 Sep 26;600(1-2):122-8. doi: 10.1016/j.aca.2007.02.021. Epub 2007 Feb 20.
The presented paper deals with a new methodology for direct determination of propranolol in human plasma. The methodology described is based on sequential injection analysis technique (SIA) coupled with solid phase extraction (SPE) column based on restricted access materials (RAM). Special RAM column containing 30 microm polymeric material-N-vinylacetamide copolymer was integrated into the sequential injection manifold. SIA-RAM system was used for selective retention of propranolol, while the plasma matrix components were eluted with two weak organic solutions to waste. Due to the acid-basic and polarity properties of propranolol molecule and principles of reversed-phase chromatography, it was possible to retain propranolol on the N-vinylacetamide copolymer sorbent (Shodex MSpak PK-2A 30 microm (2 mm x 10 mm)). Centrifuged plasma samples were aspirated into the system and loaded onto the column using acetonitrile-water (5:95, v/v), pH 11.00, adjusted by triethylamine. The analyte was retained on the column while proteins contained in the sample were removed to waste. Interfering endogenous substances complicating detection were washed out by acetonitrile-water (15:85), pH 11.00 in the next step. The extracted analyte was eluted by means of tetrahydrofuran-water (25:75), pH 11.00 to the fluorescence detector (emission filter 385 nm). The whole procedure comprising sample pre-treatment, analyte detection and column reconditioning took about 15 min. The recoveries of propranolol from undiluted plasma were in the range 96.2-97.8% for three concentration levels of analyte. The proposed SIA-RAM method has been applied for direct determination of propranolol in human plasma.
本文介绍了一种直接测定人血浆中普萘洛尔的新方法。所描述的方法基于顺序注射分析技术（SIA）与基于限进材料（RAM）的固相萃取（SPE）柱相结合。将含有30微米聚合材料-N-乙烯基乙酰胺共聚物的特殊RAM柱集成到顺序注射流路中。SIA-RAM系统用于选择性保留普萘洛尔，而血浆基质成分则用两种弱有机溶液洗脱至废液中。由于普萘洛尔分子的酸碱和极性性质以及反相色谱原理，有可能将普萘洛尔保留在N-乙烯基乙酰胺共聚物吸附剂（Shodex MSpak PK-2A 30微米（2毫米×10毫米））上。将离心后的血浆样品吸入系统，用乙腈-水（5:95，v/v），pH 11.00（用三乙胺调节）加载到柱上。分析物保留在柱上，同时样品中含有的蛋白质被排至废液中。下一步用乙腈-水（15:85），pH 11.00冲洗掉干扰检测的内源性物质。提取的分析物用四氢呋喃-水（25:75），pH 11.00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，耗时约15分钟。对于三种分析物浓度水平，普萘洛尔从未稀释血浆中的回收率在96.2-97.8%范围内。所提出的SIA-RAM方法已用于直接测定人血浆中的普萘洛尔。
