Native fluorescent detection with sequential injection chromatography for doping control analysis

Abstract

BackgroundSequential 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.ResultsAML 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.ConclusionsThe 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.

Spectrophotometric Assay Methods for Determination of Amiloride: A Charge-Transfer Reaction Approach with 2,3-Dichloro-5,6-Dicyano-1,4-Benzoquinone and p-Chloranilic Acid Reagents

“…The proposed methods basically relay on the formation of stable charge transfer complexes [11] . High Performance Liquid Chromatography (HPLC) [12][13][14][15][16][17] , some of the method uses the chemometrics in conjunction with UV-spectrophotometer [14][15][16]24] , methods uses spectrofluorimeter [17][18][19][20][21] , a method with Capillary Zone Electrophoresis [22,23] , a to assess the AMD in formulations and biological samples. However, these procedures are connected with sophistication, expertise, extraction, and more expensive than other methods.…”

Computational study of amiloride – a WADA banned molecule

“…First, they can lead to significantly faster weight loss. This refer to judo, karate and other martial arts, where category classifications are made according to weight (Idris & Alnajjar 2013). Secondly, since they increase the urine flow, they can practically cover other forbidden substances consumed to achieve better sport results (Pereira et al 2016).…”

Flow-Batch Sample Preparation for Fractionation of the Stress Signaling Phytohormone Salicylic Acid in Fresh Leaves

“…Sequential injection chromatography (SIC) combines sequential injection analysis (SIA) with monolithic or fused-core particles columns operating at relatively low pressures to promote fast and cost-effective chromatographic separations, minimizing eluent volumes and waste amounts [ 12 , 13 ]. Despite the lower chromatographic efficiency compared to HPLC, successful separations have been achieved for a diversity of applications, including determination of pharmaceutical compounds in medicines [ 12 ] and body fluids [ 14 ], contaminants in food [ 13 ], and pesticides in natural waters [ 15 ]. Barrientos et al [ 16 ] successfully applied SIC with fluorimetric detection for determination of free SA on plant leaves after batchwise microwave-assisted extraction.…”