Development of Sarcosine Quantification in Urine Based on Enzyme-coupled Colorimetric Method for Prostate Cancer Diagnosis

Overview

Journal EXCLI J

Specialty Biology

Date 2018 Jul 24

PMID 30034310

Citations 5

Authors

Vichanan Yamkamon

Benjarong Phakdee

Sakda Yainoy

Thummaruk Suksrichawalit

Tararat Tatanandana

Premsant Sangkum

Warawan Eiamphungporn

Affiliations

Soon will be listed here.

Abstract

An enzyme-coupled colorimetric assay for quantification of urinary sarcosine was developed. The proposed method is a specific reaction based on hydrogen peroxide (HO) formation via sarcosine oxidase (SOX). The liberated HO 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.

Citing Articles

Nurturing the marriages of urinary liquid biopsies and nano-diagnostics for precision urinalysis of prostate cancer.

Liao C, Wu Z, Lin C, Chen X, Zou Y, Zhao W Smart Med. 2024; 2(1):e20220020.

PMID: 39188554 PMC: 11236013. DOI: 10.1002/SMMD.20220020.

Integrating Microfluidics and Electronics in Point-of-Care Diagnostics: Current and Future Challenges.

Annese V, Hu C Micromachines (Basel). 2022; 13(11).

PMID: 36363944 PMC: 9699090. DOI: 10.3390/mi13111923.

Circulating metabolite biomarkers: a game changer in the human prostate cancer diagnosis.

Krishnan S, Kanthaje S, Punchappady D, Mujeeburahiman M, Ratnacaram C J Cancer Res Clin Oncol. 2022; 149(3):951-967.

PMID: 35764700 DOI: 10.1007/s00432-022-04113-y.

Simultaneous determination of sarcosine and its related metabolites by gas chromatography-tandem mass spectrometry for prostate cancer diagnosis.

Yamkamon V, Yee P, Yainoi S, Eiamphungporn W, Suksrichavalit T EXCLI J. 2018; 17:965-979.

PMID: 30564076 PMC: 6295632. DOI: 10.17179/excli2018-1352.

A Rapid Method for the Detection of Sarcosine Using SPIONs/Au/CS/SOX/NPs for Prostate Cancer Sensing.

Uhlirova D, Stankova M, Docekalova M, Hosnedlova B, Kepinska M, Ruttkay-Nedecky B Int J Mol Sci. 2018; 19(12).

PMID: 30467297 PMC: 6320840. DOI: 10.3390/ijms19123722.

References

MAEHLY A, Chance B . The assay of catalases and peroxidases. Methods Biochem Anal. 1954; 1:357-424. DOI: 10.1002/9780470110171.ch14. View

Jiang Y, Cheng X, Wang C, Ma Y . Quantitative determination of sarcosine and related compounds in urinary samples by liquid chromatography with tandem mass spectrometry. Anal Chem. 2010; 82(21):9022-7. DOI: 10.1021/ac1019914. View

Shamsipur M, Naseri M, Babri M . Quantification of candidate prostate cancer metabolite biomarkers in urine using dispersive derivatization liquid-liquid microextraction followed by gas and liquid chromatography-mass spectrometry. J Pharm Biomed Anal. 2013; 81-82:65-75. DOI: 10.1016/j.jpba.2013.03.019. View

Yao Y, Zhang C . A novel screen-printed microfluidic paper-based electrochemical device for detection of glucose and uric acid in urine. Biomed Microdevices. 2016; 18(5):92. DOI: 10.1007/s10544-016-0115-6. View

Kinoshita T, Hiraga Y . A fluorophotometric determination of serum creatinine and creatine using a creatinineamidohydrolase-creatineamidinohydrolase-sarcosine oxidase-peroxidase system and diacetyldichlorofluorescin. Chem Pharm Bull (Tokyo). 1980; 28(12):3501-6. DOI: 10.1248/cpb.28.3501. View

Meyer T, Fox S, Issaq H, Xu X, Chu L, Veenstra T . A reproducible and high-throughput HPLC/MS method to separate sarcosine from α- and β-alanine and to quantify sarcosine in human serum and urine. Anal Chem. 2011; 83(14):5735-40. DOI: 10.1021/ac201003r. View

Valenti G, Rampazzo E, Biavardi E, Villani E, Fracasso G, Marcaccio M . An electrochemiluminescence-supramolecular approach to sarcosine detection for early diagnosis of prostate cancer. Faraday Discuss. 2015; 185:299-309. DOI: 10.1039/c5fd00096c. View

Roobol M, Zappa M, Maattanen L, Ciatto S . The value of different screening tests in predicting prostate biopsy outcome in screening for prostate cancer data from a multicenter study (ERSPC). Prostate. 2006; 67(4):439-46. DOI: 10.1002/pros.20545. View

Ankerst D, Thompson I . Sensitivity and specificity of prostate-specific antigen for prostate cancer detection with high rates of biopsy verification. Arch Ital Urol Androl. 2007; 78(4):125-9. View

10.

Cao D, Ye D, Zhu Y, Zhang H, Wang Y, Yao X . Efforts to resolve the contradictions in early diagnosis of prostate cancer: a comparison of different algorithms of sarcosine in urine. Prostate Cancer Prostatic Dis. 2011; 14(2):166-72. DOI: 10.1038/pcan.2011.2. View

11.

Tiwari G, Tiwari R . Bioanalytical method validation: An updated review. Pharm Methods. 2013; 1(1):25-38. PMC: 3658022. DOI: 10.4103/2229-4708.72226. View

12.

Votyakova T, Reynolds I . Detection of hydrogen peroxide with Amplex Red: interference by NADH and reduced glutathione auto-oxidation. Arch Biochem Biophys. 2004; 431(1):138-44. DOI: 10.1016/j.abb.2004.07.025. View

13.

Cary K, Cooperberg M . Biomarkers in prostate cancer surveillance and screening: past, present, and future. Ther Adv Urol. 2013; 5(6):318-29. PMC: 3825107. DOI: 10.1177/1756287213495915. View

14.

Fossati P, PRENCIPE L, Berti G . Enzymic creatinine assay: a new colorimetric method based on hydrogen peroxide measurement. Clin Chem. 1983; 29(8):1494-6. View

15.

Martinez-Lozano P, Rus J . Separation of isomers L-alanine and sarcosine in urine by electrospray ionization and tandem differential mobility analysis-mass spectrometry. J Am Soc Mass Spectrom. 2010; 21(7):1129-32. DOI: 10.1016/j.jasms.2010.02.021. View

16.

Sreekumar A, Poisson L, Rajendiran T, Khan A, Cao Q, Yu J . Metabolomic profiles delineate potential role for sarcosine in prostate cancer progression. Nature. 2009; 457(7231):910-4. PMC: 2724746. DOI: 10.1038/nature07762. View

17.

Zhou M, Diwu Z, Haugland R . A stable nonfluorescent derivative of resorufin for the fluorometric determination of trace hydrogen peroxide: applications in detecting the activity of phagocyte NADPH oxidase and other oxidases. Anal Biochem. 1997; 253(2):162-8. DOI: 10.1006/abio.1997.2391. View

18.

Perez Galende P, Manzano Munoz T, Roig M, Garcia de Maria C . Use of crude extract of lentil plant (Lens culinaris Medikus) in peroxidase-based analyses: fast kinetic determination of hydrogen peroxide and sarcosine in urine. Anal Bioanal Chem. 2012; 404(8):2377-85. DOI: 10.1007/s00216-012-6343-6. View

19.

Rebelo T, Pereira C, Sales M, Noronha J, Costa-Rodrigues J, Silva F . Sarcosine oxidase composite screen-printed electrode for sarcosine determination in biological samples. Anal Chim Acta. 2014; 850:26-32. DOI: 10.1016/j.aca.2014.08.005. View

20.

Kayamori Y, Katayama Y, Urata T . Nonenzymatic elimination of ascorbic acid in clinical samples. Clin Biochem. 2000; 33(1):25-9. DOI: 10.1016/s0009-9120(99)00082-x. View