Determination of catecholamine sulfoconjugate isomers in normal human urine by use of high-performance liquid chromatography with a photochemical detector.
A method for the determination of catecholamine sulfoconjugate isomers (CA-S) in urine was developed. The photo-induced fluorogenic reaction of CA-S with ethylenediamine reported previously was applied to the postcolumn labeling of HPLC for sensitive and selective detection. Special equipment for the reaction was made with a uv-irradiation lamp and a reaction coil of Teflon tubing inside a temperature-controlled reaction box. Lower determination limits of this system were 1 to 2 pmol. Urine samples pretreated with small ion-exchange resin columns were subjected to HPLC. Peaks corresponding to CA-S were identified quantitatively by two different separation methods. Thus, all six CA-S were first detected in the urine of normal individuals. The excretion rates of dopamine 3-sulfate, dopamine 4-sulfate, norepinephrine 3-sulfate, norepinephrine 4-sulfate, epinephrine 3-sulfate, and epinephrine 4-sulfate were 420 +/- 240, 98 +/- 55, 86 +/- 95, 15 +/- 14, 18 +/- 7, and 3 +/- 1 ng/min (+/- SD), respectively (n = 5).
D002395:Catecholamines / Q000302:isolation & purification / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation*; D002852:Chromatography, Ion Exchange; D005029:Ethylenediamines; D006801:Humans; D008297:Male; D010777:Photochemistry / Q000295:instrumentation*; D013431:Sulfates / Q000302:isolation & purification
D002395:Catecholamines; D005029:Ethylenediamines; D013431:Sulfates