The study aims to evaluate a standardized Sprague-Dawley (SD) rat model for postmenopausal osteoporosis (PMO) research, including the selection of animals, surgical procedures, anesthetic dosage, and supportive care for a speedy recovery, osteoporotic induction, and biochemical and DEXA scan evaluation.Midventral ovariectomy was performed by removing ovaries to simulate a postmenopausal clinical condition in SD rats.The serum markers, estradiol, osteocalcin, and tartrateresistant acid phosphatase levels in serum were estimated, and PMO status was confirmed with a DEXA scan by evaluating the whole bone mineral density and bone mineral content.The ovariectomized rat (OVX) showed a sharp loss in bone mass and mineral content and serum calcium significantly lowered from 11.21±0.85mg/dl to 6.34±0.69mg/dl, magnesium from 2.87±0.43mg/dl to 2.11±0.31mg/dl and phosphorus 4.03±0.58mg/dl to 2.73±0.87mg/dl respectively.The OVX rats showed a significant reduction in osteocalcin and estradiol values of 1.08±0.68ng/ml and 67.28±11.46pg/ml at the end of 6 months, respectively.In OVX animals, the total bone mineral density obtained by DEXA scan was 0.164 ±0.004 g/cm 2 compared to sham control (0.183 ±0.006 g/cm 2 ).Midventral ovariectomy can be considered an accepted surgical procedure to create an animal model for understanding PMO-associated bone loss.Serum estradiol evaluation along with osteocalcin, calcium, and DEXA scan standardized the status of PMO for detailed study.