Multicommutated flow analysis system based on fluorescence microdetectors for simultaneous determination of phosphate...

A bianalyte multicommutated flow analysis (MCFA) system allowing simultaneous determination of calcium and phosphate ions is developed. The detection of ions is based on measurements of fluorescence of calcein and rhodamine B, respectively. For such measurements performed under flow analysis conditions two dedicated detectors made of three integrated light emitting diodes each and operating according to fluorometric paired emitter detector diode (FPEDD) principle, have been used. The developed 2FPEDD-MCFA system has been applied for serum analysis. The system provides low detection limits (16 µM and 3.6 µM for calcium and phosphate ions, respectively) and therefore small consumption of sample (8 µL) for analysis. The throughput of the system allows about 20 bianalyte determinations per hour. The presented system was validated using human serum samples. The analysis results show good correlation with those from the clinical laboratory obtained using reference photometric methods. The results of real samples analysis show that it is necessary to determine both calcium and phosphate ions simultaneously because their levels is serum are rather weakly correlated.