PHOTOMETRIC DETECTION OF AMINO ACID SARCOSINE USING ITS HYDROLYSIS BY SARCOSINE OXIDASE AND AMPLEX RED FLUOROPHORE

1 STANKOVA Martina
Co-authors:
1 VSETICKOVA Michaela 1 TOTHOVA Zuzana 1 UHLIROVA Dagmar 2 RUTTKAY-NEDECKY Branislav 3 KEPINSKA Marta 1 RUZICKA Josef 3 MILNEROWICZ Halina 4 EIAMPHUNGPORN Warawan 1,2,3 KIZEK Rene
Institutions:
1 Department of Research and Development, Prevention Medicals s.r.o., Tovarni 342, 742 13; Studenka-Butovice, Czech Republic, stankova@preventionmedicals.cz
2 Department of Human Pharmacology and Toxicology, University of Veterinary and Pharmaceutical Sciences Brno, Palackého třída 1946/1, 612 42 Brno, Czech Republic, kizek@sci.muni.cz
3 Department of Biomedical and Environmental Analyses, Faculty of Pharmacy with Division of Laboratory Diagnostics, Wroclaw Medical University, Borowska 211, 50-556 Wroclaw, Poland, zalewska.m@gmail.com
4 Department of Clinical Microbiology and Applied Technology, Faculty of Medical Technology, Mahidol University, 999 Phutthamonthon 4 Road, Salaya, Nakhon Pathom 73170. Thailand, warawan.eia@mahidol.ac.th
Conference:
11th International Conference on Nanomaterials - Research & Application, Hotel Voronez I, Brno, Czech Republic, EU, October 16th - 18th 2019
Proceedings:
Proceedings 11th International Conference on Nanomaterials - Research & Application
Pages:
358-363
ISBN:
978-80-87294-95-6
ISSN:
2694-930X
Published:
1st April 2020
Proceedings of the conference have been sent to Web of Science and Scopus for evaluation and potential indexing.
Metrics:
14 views / 4 downloads
Abstract

Prostate cancer is the most common malignant tumor in men, whose incidence is significantly different geographically and increases with age. For rapid diagnostics, new tumor markers with higher prognostic relevance are still being sought. A very promising candidate molecule is the amino acid sarcosine. The aim of this work was to design a highly sensitive photometric detection system for the sarcosine determination. An original methodology for determining the amino acid sarcosine using Trinder's reaction with sarcosine oxidase and Amplex Red fluorophore has been proposed. To improve the enzymatic reaction sarcosine oxidase and horseradish peroxidase were immobilised on superparamagnetic irone oxide nanoparticles (SPIONs). Absorbance dependence on sarcosine concentration read off at the wavelength of 572 nm was linear over a total range of 0 - 1000 μM (r ≥ 0.99) with limit of detection (LOD) of 2 μM and limit of quantification (LOQ) of 6 μM. The suggested procedure allows to analyze the content of sarcosine in real urine specimens. The proposed method is capable of detecting sarcosine at micromolar concentrations.

Keywords: Sarcosine; prostate cancer; photometric detection; red amplex; sarcosine oxidase
Scroll to Top