Introduction: In women with suspected urinary tract infection (UTI), a non-contaminated voided specimen is considered important for valid urinalysis and culture results. We assess whether midstream parted-labia catch (MSPC) instructions were provided by nurses, understood, and performed correctly, according to the patient. Methods: We conducted a cross-sectional survey of English- and Spanish-speaking female patients submitting voided urine samples for urinalysis for suspected UTI. The survey was conducted in a public teaching hospital emergency department (ED) from June to December 2010, beginning 2 months after development and dissemination of a nursing MSPC instructions protocol. Research assistants administered the survey within 2 hours of urine collection. Nurses were unaware of the study purpose. Results: Of 129 patients approached, 74 (57%) consented and were included in the analysis. Median age was 35; 44% were Latino. Regarding instructions from nurses, patients reported the following: 45 (61%; 95% CI 50–72%) received any instructions; of whom 37 (82%; 95% CI 71–93%) understood them completely. Sixteen (36%; 95% CI 22–51%) were instructed to collect midstream; and 7 (16%; 95% CI 6–29%) to part the labia. Regardless of receiving or understanding instructions, 33 (45%; 95% CI 33–57%) reported actually collecting midstream, and 11 (15%, 95% CI 8–25%) parting the labia. Conclusion: In this ED, instructions for MSPC urine collection frequently were not given, despite a nursing protocol, and patients rarely performed the essential steps. An evidence-based approach to urine testing in the ED that considers urine collection technique, is needed.