Background Random urine protein-to-creatinine (PCR) and albumin-to-creatinine (ACR) ratios have been proposed as alternatives to 24 h urine measurements to simplify sample collection and overcome errors. The aim of this study was to examine the ability of PCR and ACR to predict urinary 24 h protein and albumin loss, respectively, in patients with kidney disease, and determine the most appropriate time of collection. Methods Eighty-three patients were recruited from a renal outpatient clinic. In a 24 h period, each collected an early-morning urine (EMU), second and third voids, and the remaining urine passed that day. PCR and ACR were determined in random urines and compared with the 24 h loss of protein and albumin, respectively. Results For all patients, median (range) 24 h urine protein and albumin losses were 220 (30–15600) and 60 (<8–10,557) mg, respectively. Ratios derived from each of three random urines correlated well with 24 h protein or albumin loss (Spearman's r s > 0.87, P < 0.0001). Receiver operator characteristic (ROC) curve analysis showed PCR accurately predicted both an abnormal 24 h urine protein ≥150 mg/24 h (areas under curves [AUC] 0.90–0.92) and significant proteinuria above 300 mg/24 h (AUC between 0.97 and 1.00). ACR accurately predicted both an abnormal 24 h urine albumin ≥30 mg/24 h (AUC 0.98 to 0.99) and frank albuminuria at ≥300 mg/24 h or ≥700 mg/24 h (AUC between 0.99 and 1.00). EMU and random urines performed equally well in predicting proteinuria and albuminuria from PCR and ACR, respectively. Conclusions By careful choice of cut-offs, both PCR and ACR can be used in patients with kidney disease to rule in or rule out abnormal 24 h losses of protein and albumin. EMU and, importantly, random samples can be used as surrogates for 24 h urine collections.