Objective: To determine if there is correlation between nighttime 12-hour and traditional 24-hour urine collection in regard to chemistry values and the supersaturations of calcium oxalate, calcium phosphate, and uric acid for the metabolic evaluation of nephrolithiasis. Materials and Methods: Ninety-five patients were prospectively enrolled from 2013 to 2015. Patients >18 years of age who presented to a tertiary stone clinic and who would normally be counseled for 24-hour urine collection were eligible for the study. Participants completed 24-hour urine collections twice, with each divided into 2 separate 12-hour collections. Day-time collection began after the first morning void and continued for 12 hours. The night collection proceeded for the next 12 hours through the first morning void. Results: Forty-nine 24-hour samples from 35 patients met inclusion criteria and were included in the analysis. Overall, there was strong correlation between the night 12-hour and the 24-hour urine collections with R2 ranging from 0.76 for pH to 0.96 for Citrate. In our analysis of variability, the nighttime 12-hour collection differed from the 24-hour collection by 30% in 1-9 patients (2.0%-18.4%) based on individual chemistry value. Diagnosis of underlying metabolic abnormalities was concordant in 92% of patients. Conclusion: A 12-hour nighttime collection has strong correlation with 24-hour urine collection. As such, simplifying the metabolic evaluation to a 12-hour overnight collection may be feasible—improving compliance and decreasing patient burden.