ABSTRACT
In this paper an ultrasensitive method to determine toxicity due to pesticides in a glass lab-on-a-chip by means of enzymatic inhibition of acetylcholinesterase immobilised on magnetic beads is described. The reproducible insertion of a controlled amount of enzyme-coupled magnetic beads inside the chip channel and their immobilisation in a capture region with the aid of a magnetic field has been optimised. This procedure enables the easy renewal of the biosensing material after each determination in a highly reproducible manner. Several operational parameters such as the working potential for the selective detection of thiocholine (TCh) on a platinum disc electrode, the TCh detection reproducibility and sensitivity, the electroosmotic flow driving voltage and the inhibition time were also evaluated or optimised. The detection of carbofuran (one of the most toxic carbamate pesticides) has been achieved down to the nanomolar level.
Subject(s)
Carbofuran/analysis , Magnetics , Microchip Analytical Procedures/methods , Pesticides/analysis , Acetylcholinesterase/metabolism , Biosensing Techniques/methods , Carbofuran/chemistry , Cholinesterase Inhibitors/chemistry , Electrodes , Enzymes, Immobilized/antagonists & inhibitors , Platinum/chemistry , Reproducibility of Results , Sensitivity and Specificity , Thiocholine/chemistryABSTRACT
Among the several fabrication techniques used to construct microflow systems, the low-temperature cofired ceramics (LTCC) technology, taking advantage of its multilayer approach, is one of the most versatile ones. It permits the integration of several unitary operations of an analytical process in a modular or monolithic way. Moreover, due to its perfect compatibility with screen-printing techniques, it also permits the integration of electronic components used to control the whole system setup. In this work the design, construction, and evaluation of a miniaturized analyzer for pesticide determination that integrates a pretreatment stage, based on two mixers or reactors, and an amperometric detection system to measure the product of an enzymatic inhibition reaction are presented. The detection system was monolithically integrated in the microfluidic platform, and it consisted of a screen-printed reference electrode and two platinum sheets, acting as auxiliary and working electrodes, which were embedded within the ceramic structure. The miniaturized system was characterized and successfully evaluated by determining carbofuran at the nanomolar level.
