A simple, precise and rapid reversed phase HPLC method was developed for the estimation of urinary crosslink Pyridinoline (PYD) and Deoxypyridinoline (DPD) in premenopausal and postmenopausal women. The method was carried out on a C-18 reverse phase column (LichroCART, 150 X 4mm, 5μm; Merck) using 2 mM Heptafluorobutyric acid: Acetonitrile (95:5) as the mobile phase and detection was carried out using Water32 2475 multi λ fluorescence detector (Excitation λ 295 nm, Emission λ 395 nm). The retention times for PYD and DPD were found to be 10.7 ± 0.2 minutes and 11.7 ± 0.2 minutes respectively. The % recovery for PYD and DPD in urine matrix was found to be 102.89 % and 108.33 % respectively. Form the study it was found that the postmenopausal women are at a higher risk of bone resorption due to high bone fragility as compared to the premenopausal women.