Colorimetric Assay Of Arginine And Spermine In Biofluids By Uv-Vis Spectrometry

INTRODUCTION

Arginine (Arg) and spermine are vital biomolecules found in biofluids, playing essential roles in cell functions, wound healing, and protein synthesis. Accurate detection and quantification of these analytes are crucial for understanding cellular processes and diagnosing diseases. Traditional analysis methods are complex and costly. So, a new colorimetric assay has been developed using 6-aza-2-thiothymine-functionalized silver nanoparticles (6-ATT-AgNPs) for precise detection of arginine and spermine in biofluids. This application note highlights the detailed methodology, sensing mechanism, experimental setup, and implications of this innovative assay in bioanalytical chemistry for easy sharing and understanding by all.

EXPERIMENTAL METHODS

The experimental procedures planned a colorimetric assay for quantifying arginine and spermine in biofluids. Calibration standards were prepared, and samples were analyzed using UV-Vis spectrometry to understand nanoparticle interactions. UV-Vis spectrometry, a powerful analytical technique capable of providing detailed insights into the absorption characteristics of the nanoparticles and their interactions with the analytes. The data obtained from the spectrometric analysis were then subjected to rigorous interpretation, enabling the extraction of meaningful information regarding the detection and quantification of arginine and spermine.

The colorimetric assay relied on the aggregation of 6-ATT-AgNPs through noncovalent interactions with the analytes, resulting in significant changes in fluorescence intensity and absorption characteristics. The sensor effectively detected arginine and spermine, ensuring reproducibility and enhancing assay effectiveness in bioanalytical applications.

Figure 1 demonstrates the colorimetric sensitivity of 6-ATT-AgNPs towards arginine (Arg) and spermine was evaluated through UV-visible absorption spectra and digital photographs. The absorption spectra of 6-ATT-AgNPs were altered with increasing concentrations of Arg (5.0–1000 uM) and spermine (2.5–1000 uM), showing distinct changes corresponding to the presence of each analyte.  

The color of 6-ATT-AgNPs shifted gradually with the presence of Arg and spermine, indicating an increased aggregation rate with higher concentrations. The linear correlation between the intensity changes and the analyte concentrations further validated the sensor's performance and accuracy in detecting arginine and spermine in biofluids.

Figure 1. (a) Absorbance spectra of 6-ATT-AgNPs with increasing Arg concentration (5.0-1000 uM). (b) Absorbance spectra of 6-ATT-AgNPs with increasing concentration of spermine (2.5-1000 uM). Insets show the color of 6-ATT-AgNPs with increasing concentrations of both analytes.

APPLICATIONS AND IMPLICATIONS

The colorimetric assay with 6-ATT-AgNPs presents a valuable tool for readers seeking an improved cancer diagnosis. By quantifying arginine and spermine levels accurately, this assay offers a fast, cost-effective, and reliable method for detecting lung, liver, prostate, breast, and colorectal cancers. The integration of fluorescence spectroscopy and UV-Vis spectrometry enhances biomarker detection, providing readers with advanced insights into cancer diagnosis and treatment. This innovative approach in bioanalytical sciences holds promise for improving cancer detection and understanding biomolecular interactions in biofluids.

recommended EQUIPMENT

In this study, the absorption characteristics play a key role in developing a straightforward colorimetric method for detecting Arg and spermine in biofluids. The absorption spectra were analyzed using the Maya 2000 Pro UV-Vis spectrometer, renowned for its exceptional sensitivity in absorbance measurements. The Maya 2000 Pro is an ideal instrument for applications demanding high sensitivity and reliable UV-Vis response. Its versatility extends to various analyses, including low light-level fluorescence, absorbance, transmission, and emission studies.

Figure 2. Maya 2000 Pro Spectrometer Series

REFERENCE

Kailasa, Suresh Kumar, et al. "Independent spectral characteristics of functionalized silver nanoparticles for colorimetric assay of arginine and spermine in biofluids." New Journal of Chemistry 43.43 (2019): 17069-17077.