Paper-Based In-Situ Gold Nanoparticle Synthesis for Colorimetric, Non-Enzymatic Glucose Level Determination.

Tomás Pinheiro,Maria J Oliveira,M Paula Macedo,Ana C Marques,João Ferrão,Pedro M F J Costa,Nitin M Batra,Hugo Águas,Elvira Fortunato,Rodrigo Martins

doi:10.3390/nano10102027

Abstract

Due to its properties, paper represents an alternative to perform point-of-care tests for colorimetric determination of glucose levels, providing simple, rapid, and inexpensive means of diagnosis. In this work, we report the development of a novel, rapid, disposable, inexpensive, enzyme-free, and colorimetric paper-based assay for glucose level determination. This sensing strategy is based on the synthesis of gold nanoparticles (AuNPs) by reduction of a gold salt precursor, in which glucose acts simultaneously as reducing and capping agent. This leads to a direct measurement of glucose without any enzymes or depending on the detection of intermediate products as in conventional enzymatic colorimetric methods. Firstly, we modelled the synthesis reaction of AuNPs to determine the optical, morphological, and kinetic properties and their manipulation for glucose sensing, by determining the influence of each of the reaction precursors towards the produced AuNPs, providing a guide for the manipulation of nucleation and growth. The adaptation of this synthesis into the developed paper platform was tested and calibrated using different standard solutions with physiological concentrations of glucose. The response of the colorimetric signals obtained with this paper-based platform showed a linear behavior until 20 mM, required for glycemic control in diabetes, using the Red × Value/Grey feature combination as a calibration metric, to describe the variations in color intensity and hue in the spot test zone. The colorimetric sensor revealed a detection limit of 0.65 mM, depending on calibration metric and sensitivity of 0.013 AU/mM for a linear sensitivity range from 1.25 to 20 mM, with high specificity for the determination of glucose in complex standards with other common reducing interferents and human serum.

Highlights

Glucose is the primary energy source of cells and acts as a metabolic intermediate, being transported through the bloodstream of organisms [1,2]
This leads to formation of Au seeds in solution, whose size corresponds to a color shift depending on glucose concentration in the system, as previously reported [36,41,42]
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Summary

Introduction

Glucose is the primary energy source of cells and acts as a metabolic intermediate, being transported through the bloodstream of organisms [1,2]. The normal levels of glucose in human blood range from 3.8 to 6.9 mM (68–124 mg/dL), approximately, these levels may change during the day, being lower upon fasting and reaching a peak in postprandial states [3]. Both high and low glycemic levels are detrimental for health status, for example, a level below 2.8 mM (50 mg/dL) after fasting or following exercise is considered an hypoglycemic event [4]. The incidence of glucose levels outside the normal range may indicate the presence of health conditions, such as diabetes mellitus [1]. Blood glucose concentration should be strictly controlled and below 10 mM (180 mg/dL) according to the American Diabetes Association [6], otherwise it can lead to serious complications including diabetic retinopathy, kidney failure, strokes, heart attacks, fatty liver disease, high blood pressure, blindness, and coma [7]

Methods

Results

Conclusion