[Analysis of the pyridinoline and deoxypyridinoline in urine by reversed-phase high performance liquid chromatography (RP-HPLC)].
A method for the simultaneous and rapid determination of pyridinoline and deoxypyridinoline in urine using RP-HPLC has been established. After hydrolysis for 18 hours at 107 degrees C in 6 mol/L HCl, the urine sample was prefractionated by partition chromatography using a column packed with cellulose CF1. The appropriate fractions were freeze-dried. The samples were analyzed by HPLC with a 5 microm Spherisorb C18 column, a mobile phase of MeOH/H2O containing 0.1% heptafluorobutyric acid at a rate of 1.2 mL/min and fluorescence detector monitoring at lambda(ex) 290 nm and lambda(em) 400 nm. The limit of a detection for PYD was 10 nmol/L and DPD 7 nmol/L. The recovery of PYD was 91.5% and that of DPD was 106.1%. Within day CV for the two compounds were 1.39% and 0.16% respectively and day-to-day CV 3.71% and 1.32% respectively.
Weng
J
J
Research Unit of Endocrinology, First Affiliated Hospital, Sun Yat-sen University of Medical Sciences, Guangzhou, 510080.
Liao
Y
Y
Yu
B
B
D000596:Amino Acids / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D006801:Humans
D000596:Amino Acids; C015484:pyridinoline; C036020:deoxypyridinoline