Development of sarcosine quantification in urine based on enzyme-coupled colorimetric method for prostate cancer diagnosis

Authors

  • Vichanan Yamkamon Department of Clinical Microscopy, Faculty of Medical Technology, Mahidol University, Bangkok 10700, Thailand
  • Benjarong Phakdee Department of Clinical Microbiology and Applied Technology, Faculty of Medical Technology, Mahidol University, Bangkok 10700, Thailand
  • Sakda Yainoy Department of Clinical Microbiology and Applied Technology, Faculty of Medical Technology, Mahidol University, Bangkok 10700, Thailand
  • Thummaruk Suksrichawalit Center of Data Mining and Biomedical Informatics, Faculty of Medical Technology, Mahidol University, Bangkok 10700, Thailand
  • Tararat Tatanandana Department of Clinical Chemistry, Faculty of Medical Technology, Mahidol University, Bangkok 10700, Thailand
  • Premsant Sangkum Department of Surgery, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok 10400, Thailand
  • Warawan Eiamphungporn Department of Clinical Microbiology and Applied Technology, Faculty of Medical Technology, Mahidol University, Bangkok 10700, Thailand

DOI:

https://doi.org/10.17179/excli2018-1245

Keywords:

colorimetric assay, sarcosine, prostate cancer, sarcosine oxidase, urine

Abstract

An enzyme-coupled colorimetric assay for quantification of urinary sarcosine was developed. The proposed method is a specific reaction based on hydrogen peroxide (H2O2) formation via sarcosine oxidase (SOX). The liberated H2O2 reacts with Amplex Red in the presence of horseradish peroxidase (HRP) to produce the red-fluorescent oxidation product, resorufin, which can be measured spectrophotometrically (OD570). The method was performed in the 96-well microtiter plate. Reaction conditions, such as pH and reaction time were optimized. At the optimum conditions, the limit of detection (LOD) and quantification (LOQ) were found to be 0.7 and 1 µM, respectively. A good linearity was revealed with a coefficient of 0.990. The assay showed no significant interference from ascorbic acid, glucose and bilirubin. In addition, it is extremely specific for sarcosine rather than other amino acids. The determination of sarcosine in human urine displayed high accuracy and good reproducibility. This method is promising to differentiate prostate cancer patients from healthy subjects according to urinary sarcosine level. Altogether, this study provides a rapid, simple and specific tool to determine urinary sarcosine which could be useful for prostate cancer diagnosis.

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Published

2018-05-17

How to Cite

Yamkamon, V., Phakdee, B., Yainoy, S., Suksrichawalit, T., Tatanandana, T., Sangkum, P., & Eiamphungporn, W. (2018). Development of sarcosine quantification in urine based on enzyme-coupled colorimetric method for prostate cancer diagnosis. EXCLI Journal, 17, 467–478. https://doi.org/10.17179/excli2018-1245

Issue

Vol. 17 (2018)

Section

Original articles

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