Determination of neopterin, kynurenine, tryptophan and creatinine in human serum by high throuput HPLC
Abstract
A new HPLC method for simultaneous determination of neopterin, creatinine, kynurenine and tryptophan in human serum was developed and validated. Monolithic stationary phase's technology (two monolithic columns RP-18e were connected with guard monolithic cartridge 4.6 mm x 50 mm + 3.0 mm x 100 mm and 4.6 x 10 mm) and special auto sampler for micro titration plates (samples are storage in dark cooled place protected against evaporation) were combined with easy sample preparation step.
As mobile phase 15 mmol/L phosphate buffer at pH 4.50 was used. Neopterin and tryptophan were detected using fluorescent detection and kynurenine and creatinine were detected by diode-array detection.
This method may be suitable for large sequences of samples in clinical research and routine practice.