Analysis for calcium, magnesium, phosphorus, oxalate, uric acid, and creatinine in 24-h urine collections is often needed for the differential diagnosis of patients with renal calculi. Considerable attention has been given to improving the methods of analysis, but improper sample collection and processing can cause significant errors for calcium and oxalate in urine samples not treated with acid and for uric acid in urine samples not treated with base. The errors are related to the concentration of the analyte, the interval the sample is stored before analysis, and the original pH of the urine sample. We describe here a system of sequential acidification (to pH 1.5) and alkalinization (to pH 9) of 24-h urine samples, followed by heating at 56 degrees C for 10 min. This procedure allows accurate analysis for all the above analytes in the same 24-h collection of urine. We validated the sample-treatment protocol for 80 24-h urine collections.