Determination of urea using high-performance liquid chromatography with fluorescence detection after automated derivatisation with xanthydrol.
A high-performance liquid chromatography (HPLC) method for the determination of urea that incorporates automated derivatisation with xanthydrol (9H-xanthen-9-ol) is described. Unlike the classic xanthydrol approach for the determination of urea, which involves the precipitation of dixanthylurea (N,N'-di-9H-xanthen-9-ylurea), the derivatisation procedure employed in this method produces N-9H-xanthen-9-ylurea, which remains in solution and can be quantified using fluorescence detection (lambda(ex)=213 nm; lambda(em)=308 nm) after chromatographic separation from interferences. The limit of detection for urea was 5 x 10(-8) M (0.003 mg L(-1)). This method was applied to the determination of urea in human and animal urine and in wine.
Clark
Shona
S
School of Life and Environmental Sciences, Deakin University, Geelong, Victoria 3217, Australia.
Francis
Paul S
PS
Conlan
Xavier A
XA
Barnett
Neil W
NW
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*; D014508:Urea / Q000032:analysis*; D014920:Wine / Q000032:analysis; D014966:Xanthenes / Q000737:chemistry*
D014966:Xanthenes; C009690:xanthydrol; D014508:Urea