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Metabolic changes due to menopause may alter urine composition and kidney stone risk but results of prior work on this association have been mixed. We examined menopause and the risk of incident kidney stones, and changes in 24-hour urine composition in the NHS (Nurses' Health Study) II.Using multivariate adjusted Cox proportional hazards models we prospectively analyzed 108,639 NHS II participants who provided information on menopause and kidney stones. We also analyzed 24-hour urine collections from 658 participants who performed a collection while premenopausal and a repeat collection after menopause.During 22 years of followup there were 3,456 incident kidney stones. The multivariate adjusted relative risk of an incident kidney stone in postmenopausal participants compared with premenopause was 1.27 (95% CI 1.08–1.46). On stratified analysis compared with premenopause the multivariate adjusted relative risk of natural and surgically induced menopause was 1.27 (95% CI 1.09–1.48) and 1.43 (95% CI 1.19–1.73), respectively. Among the 74,505 postmenopausal participants there was a total of 1,041 incident stone events. Compared with no hormone therapy neither current nor past use was significantly associated with kidney stone risk. Compared with premenopause the postmenopausal urine collections had lower mean calcium, citrate, phosphorus and uric acid, and higher mean volume.Postmenopausal status is associated with a higher risk of incident kidney stones. Natural menopause and surgical menopause are independently associated with higher risk. There are small but significant differences in urine composition between premenopausal and postmenopausal urine collections.