No abstract

Highly sensitive label-free techniques of DNA determination are particularly interesting in relation to the present development of the DNA sensors. We show that subnanomolar concentrations (related to monomer content) of unlabeled DNA can be determined using copper solid amalgam electrodes or hanging mercury drop electrodes in the presence of copper. DNA is first treated with acid (e.g., 0.5 M perchloric acid), and the acid-released purine bases are directly determined by the cathodic stripping voltammetry. Volumes of 5-3 microL of acid-treated DNA can easily be analyzed, thus making possible the determination of picogram and subpicogram amounts of DNA corresponding to attomole and subattomole quantities of 1000-base pair DNA. Application of this determination in DNA hybridization detection is demonstrated using surface H for the hybridization (superparamagnetic beads with covalently attached DNA probe) and the mercury electrodes only for the determination of DNA selectively captured at surface H.

The preparation, activation and electrochemical pretreatment of electrodes based on nontoxic solid amalgams were described. Testing of metal solid amalgam electrodes (MeSAEs) proved their broad applicability in many respects, e.g., as to the range of working potentials and the level of background currents, well comparable with those of the hanging mercury drop electrode (HMDE). A regeneration of their surfaces before each measurements could be simply automatized using a PC-controlled system providing a reasonable repeatability of voltammetric measurements down to 3% RSD. Combination with stripping techniques at accumulation times t ac 300 s the detection limit amounted to the concentration level of 1 ppb Cu(II), Pb(II), Cd(II), Zn(II), etc. Best electrochemical properties were exhibited by the silver solid amalgam electrode (AgSAE). For example, polished AgSAE (p-AgSAE), completely free of liquid mercury, proved satisfactory even at more negative potentials enabling the determination of Zn(II), Mn(II), IO À 3 , etc. Moreover, even better repeatability of mercury meniscus modified AgSAE (m-AgSAE) was due to better quality and renewability of its surface. In many cases further testing confirmed that under appropriate conditions MeSAEs represent good, often cheaper and more users-friendly alternatives to HMDE.

This review describes recent results regarding voltammetric and amperometric determination of submicromolar concentrations of various environmentally important biologically active organic substances using nontraditional types of electrodes either in batch analysis or in flow liquid systems (especially HPLC or FIA with electrochemical detection). Attention is paid to solid amalgam electrodes (environmentally friendly alternatives to mercury electrodes), to carbon paste electrodes with easily renewable surface, to boron doped diamond film electrodes with very low noise and broad potential window, and to inexpensive solid composite electrodes with high signal-to-noise ratio, compatibility with organic solvents and easy mechanical or electrochemical pretreatment. The review concentrates on our own results in the context of the general development in the filed.