Fluorescence derivatisation of urinary corticosteroids for high-performance liquid chromatographic analysis

Simple liquid chromatography method for the rapid simultaneous determination of prednisolone and cortisol in plasma and urine using hydrophilic lipophilic balanced solid phase extraction cartridges

“…Other methods use HPLC-mass spectrometry [10][11][12] which has the disadvantage of being expensive and not available in all laboratories. Some of these methods use fluorimetric derivatisation before HPLC analysis [5,13] which is limited by the precisely controlled reaction conditions and the instability of the fluorescent analogue [9]. Many of these methods give rise to long retention times for prednisolone and cortisol of up to 20 min [4,14,15].…”

HPLC method for determination of fluorescence derivatives of cortisol, cortisone and their tetrahydro- and allo-tetrahydro-metabolites in biological fluids

“…Similarly, shorter retention times were obtained for F and E if compared to method presented by Shibata et al [36]. Resolution of the steroidic compounds was received using simple mobile phase consisted of acetonitrile and acidified water (pH 4.6) which is also much more friendly to the environment than applied earlier [36,37] organic normal phases.…”

“…The assay following the elaborated method is the least timeconsuming with comparison to other HPLC-FLD methods [36,37]. The retention times presented in the literature [37] were for A-ring metabolites of F and E about 45-50 min but the separation of F and E was not provided. Similarly, shorter retention times were obtained for F and E if compared to method presented by Shibata et al [36].…”

“…The reaction of steroids with 9-anthroyl nitrile to receive fluorescent derivatives has been reported [36][37][38]. The temperature of the reaction was found as very important in reaction selectivity, especially considering tetrahydro-steroids as THF, allo-THF and THE.…”

“…HPLC-normal phase with fluorescence detection was applied for estimating 9-anthroyl nitrile (9-AN) derivatives of F and E in plasma and urine in presence of prednisolone and prednisone [36]. 9-AN derivatives of urinary metabolites of F and E (THF, allo-THF and THE) were determined in similar HPLC system [37]. RP-HPLC method with fluorescence detection was evaluated for determination of triamcinolone (TMC) in the presence of endogenous glucocorticoids [38].…”