Monitoring potential prostate cancer biomarkers in urine by capillary electrophoresis–tandem mass spectrometry

Current prostate cancer (PCa) diagnosis based on prostate-specific antigen (PSA) has been gradually losing its credibility over the last decade due to contradictory results in published literature and clinical practice. Recently, a group of potential PCa biomarkers in urine, particularly sarcosine, was found to increase significantly as the cancer progressed to metastasis. We report a simple, robust, and reproducible CE–ESI-MS/MS method for the determination of sarcosine and other representative potential biomarkers in pooled urine. The pooled urine was obtained from 20 healthy adult volunteers between the ages of 23–30 years old. A solid phase extraction (SPE) technique was optimized for maximum recovery of sarcosine. With no derivatization step, excellent resolution between sarcosine and its isomers (α-alanine and β-alanine) was achieved. A separate non-SPE method was also developed for quantitative determination of highly concentrated urinary metabolites. CE separation was performed on a positively-charged, polyethyleneimine (PEI)-coated capillary using 0.4–2% formic acid in 50% methanol. Precision for intra- and inter-day standard addition calibration of sarcosine were found to be within 15%, whereas intra-day precisions for the rest of the metabolites varied from 0.03 to 13.4%. Acceptable intra-day and inter-day accuracies, ranging from 80 to 124%, were obtained for sarcosine and the other metabolites.

► A CE–MS/MS method for potential prostate cancer biomarkers was developed.
► Solid phase extraction was needed for low level metabolites such as sarcosine.
► The method is ready to be used for prostate cancer diagnosis and prognosis.