Simultaneous liquid chromatographic determination of creatinine and pseudouridine in bovine urine and the effect of sample pH on the analysis.
A rapid, reliable method for the simultaneous determination of creatinine and pseudouridine is described. Both analytes were detected at an optimum wavelength of detection (262 nm), considering the relative levels present in bovine urine. Cimetidine was used as the internal standard and detected at its maximum wavelength of absorption (220 nm) on a separate channel. All three compounds were eluted within 15 min, using a 10 mmol/L phosphate buffer (pH 6.8)-methanol gradient on a C18 column. Creatinine data were found to be significantly dependent upon the pH of the sample. Recoveries of both analytes were above 96%. Lowest detectable levels of creatinine and pseudouridine were 0.28 nmol and 9.0 pmol, respectively. The use of internal standard resulted in a method with high precision (standard deviation of 1.42 mmol/L and 0.027 mmol/L for creatinine and pseudouridine), yet one that was simple and rapid.
Hewavitharana
Amitha K
AK
Cooperative Research Centre for Cattle and Beef Quality, CSIRO Food Science Australia, Cannon Hill, Queensland 4170, Australia. Amitha.Hewavitharana@csiro.au
Bruce
Heather L
HL
D000818:Animals; D002417:Cattle / Q000652:urine*; D002851:Chromatography, High Pressure Liquid; D003404:Creatinine / Q000652:urine*; D006863:Hydrogen-Ion Concentration; D011560:Pseudouridine / Q000652:urine*; D012680:Sensitivity and Specificity
D011560:Pseudouridine; D003404:Creatinine