Idris Abubakr M, Alnajjar Ahmed O
Department of Chemistry, College of Science, King Khalid University, P,O, Box 9004, Abha, Saudi Arabia.
Chem Cent J. 2013 Aug 28;7(1):144. doi: 10.1186/1752-153X-7-144.
Sequential injection chromatography (SIC) is a young, ten years old, separation technique. It was proposed with the benefits of reagent-saving, rapid analysis, system miniaturization and simplicity. SIC with UV detection has proven to be efficient mostly for pharmaceutical analysis. In the current study, a stand-alone multi-wavelength fluorescence (FL) detector was coupled to an SIC system. The hyphenation was exploited for developing an SIC-FL method for the separation and quantification of amiloride (AML) and furosemide (FSM) in human urine and tablet formulation.
AML and FSM were detected using excitation maxima at 380 and 270 nm, respectively, and emission maxima at 413 and 470 nm, respectively. The separation was accomplished in less than 2.0 min into a C18 monolithic column (50 × 4.6 nm) with a mobile phase containing 25 mmol/L phosphate buffer (pH 4.0): acetonitrile: (35:65, v/v). The detection limits were found to be 12 and 470 ng/mL for AML and FSM, respectively.
The proposed SIC-FL method features satisfactory sensitivity for AML and FSM in urine samples for the minimum required performance limits recommended by the World Anti-Doping Agency, besides a downscaled consumption of reagents and high rapidity for industrial-scale analysis of pharmaceutical preparations.
顺序注射色谱法（SIC）是一种新兴的、已有十年历史的分离技术。它具有节省试剂、分析快速、系统小型化和操作简单等优点。已证明配备紫外检测的SIC在药物分析中大多是高效的。在本研究中，将一台独立的多波长荧光（FL）检测器与SIC系统联用。利用这种联用技术开发了一种SIC-FL方法，用于分离和定量人尿液及片剂制剂中的阿米洛利（AML）和呋塞米（FSM）。
分别使用380 nm和270 nm的最大激发波长以及413 nm和470 nm的最大发射波长检测AML和FSM。在不到2.0分钟的时间内，在一根C18整体柱（50×4.6 nm）上完成分离，流动相为含25 mmol/L磷酸盐缓冲液（pH 4.0）：乙腈（35:65，v/v）。发现AML和FSM的检测限分别为12 ng/mL和470 ng/mL。
所提出的SIC-FL方法除了试剂消耗减少和对药物制剂进行工业规模分析速度快之外，对于尿液样本中的AML和FSM，在世界反兴奋剂机构推荐的最低性能要求限度内具有令人满意的灵敏度。
