Analysis of urinary pseudouridine by micellar electrokinetic capillary chromatography.
We describe a capillary electrophoresis procedure, using uridine as an internal standard, for the analysis of urinary pseudouridine following solid-phase extraction. This method retains the advantages of existing chromatographic techniques but has superior resolving power and is technically less demanding. The standard curves were linear and reproducible with a detection limit of 60 fmol; chromatographic analysis was complete in under 10 min. Injection variability was < 5% and multiple independent analyses of the same urine sample for pseudouridine concentration gave coefficients of variation of < 10%. The mean (SD) urinary pseudouridine level in 18 healthy subjects was 16.1 (2.1) nmol/mumol creatinine. For a limited group of subjects where samples were taken more frequently, intra-individual variation averaged 27.5% reflecting variable excretion.
Evans
M D
MD
Division of Chemical Pathology, University of Leicester, UK.
Perrett
D
D
Lunec
J
J
Herbert
K E
KE
D000293:Adolescent; D000328:Adult; D002648:Child; D002853:Chromatography, Liquid / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005260:Female; D006801:Humans; D008297:Male; D011560:Pseudouridine / Q000652:urine*; D012016:Reference Values; D015203:Reproducibility of Results; D013048:Specimen Handling; D014529:Uridine / Q000652:urine
D011560:Pseudouridine; D014529:Uridine