Dionysios Christodouleas

Universal mobile electrochemical detector designed for use in resource-limited applications

Significance The ability to perform electrochemical testing in the field, and in resource-limited environments, and to transmit data automatically to “the cloud” can enable a broad spectrum of analyses useful for personal and public health, clinical analysis, food safety, and environmental monitoring. Although the developed world has many options for analysis and web connection, the developing world does not have broad access to either the expensive equipment necessary to perform these tests or the advanced technologies required for network connectivity. To overcome these limitations, we have developed a simple, affordable, handheld device that can perform all the most common electrochemical analyses, and transmit the results of testing to the cloud from any phone, over any network, anywhere in the world. This paper describes an inexpensive, handheld device that couples the most common forms of electrochemical analysis directly to “the cloud” using any mobile phone, for use in resource-limited settings. The device is designed to operate with a wide range of electrode formats, performs on-board mixing of samples by vibration, and transmits data over voice using audio—an approach that guarantees broad compatibility with any available mobile phone (from low-end phones to smartphones) or cellular network (second, third, and fourth generation). The electrochemical methods that we demonstrate enable quantitative, broadly applicable, and inexpensive sensing with flexibility based on a wide variety of important electroanalytical techniques (chronoamperometry, cyclic voltammetry, differential pulse voltammetry, square wave voltammetry, and potentiometry), each with different uses. Four applications demonstrate the analytical performance of the device: these involve the detection of (i) glucose in the blood for personal health, (ii) trace heavy metals (lead, cadmium, and zinc) in water for in-field environmental monitoring, (iii) sodium in urine for clinical analysis, and (iv) a malarial antigen (Plasmodium falciparum histidine-rich protein 2) for clinical research. The combination of these electrochemical capabilities in an affordable, handheld format that is compatible with any mobile phone or network worldwide guarantees that sophisticated diagnostic testing can be performed by users with a broad spectrum of needs, resources, and levels of technical expertise.

Folding analytical devices for electrochemical ELISA in hydrophobic R(H) paper.

This work describes a device for electrochemical enzyme-linked immunosorbent assay (ELISA) designed for low-resource settings and diagnostics at the point of care. The device is fabricated entirely in hydrophobic paper, produced by silanization of paper with decyl trichlorosilane, and comprises two zones separated by a central crease: an embossed microwell, on the surface of which the antigen or antibody immobilization and recognition events occur, and a detection zone where the electrodes are printed. The two zones are brought in contact by folding the device along this central crease; the analytical signal is recorded from the folded configuration. Two proof-of-concept applications, an electrochemical direct ELISA for the detection of rabbit IgG as a model antigen in buffer and an electrochemical sandwich ELISA for the detection of malarial histidine-rich protein from Plasmodium falciparum (Pf HRP2) in spiked human serum, show the versatility of this device. The limit of detection of the electrochemical sandwich ELISA for the quantification of Pf HRP2 in spiked human serum was 4 ng mL(-1) (10(2) pmol L(-1)), a value within the range of clinically relevant concentrations.

Integrating Electronics and Microfluidics on Paper

Paper microfluidics and printed electronics have developed independently, and are incompatible in many aspects. Monolithic integration of microfluidics and electronics on paper is demonstrated. This integration makes it possible to print 2D and 3D fluidic, electrofluidic, and electrical components on paper, and to fabricate devices using them.

Broadly Available Imaging Devices Enable High-Quality Low-Cost Photometry

This paper demonstrates that, for applications in resource-limited environments, expensive microplate spectrophotometers that are used in many central laboratories for parallel measurement of absorbance of samples can be replaced by photometers based on inexpensive and ubiquitous, consumer electronic devices (e.g., scanners and cell-phone cameras). Two devices, (i) a flatbed scanner operating in transmittance mode and (ii) a camera-based photometer (constructed from a cell phone camera, a planar light source, and a cardboard box), demonstrate the concept. These devices illuminate samples in microtiter plates from one side and use the RGB-based imaging sensors of the scanner/camera to measure the light transmitted to the other side. The broadband absorbance of samples (RGB-resolved absorbance) can be calculated using the RGB color values of only three pixels per microwell. Rigorous theoretical analysis establishes a well-defined relationship between the absorbance spectrum of a sample and its corresponding RGB-resolved absorbance. The linearity and precision of measurements performed with these low-cost photometers on different dyes, which absorb across the range of the visible spectrum, and chromogenic products of assays (e.g., enzymatic, ELISA) demonstrate that these low-cost photometers can be used reliably in a broad range of chemical and biochemical analyses. The ability to perform accurate measurements of absorbance on liquid samples, in parallel and at low cost, would enable testing, typically reserved for well-equipped clinics and laboratories, to be performed in circumstances where resources and expertise are limited.

Development and validation of a chemiluminogenic method for the evaluation of antioxidant activity of hydrophilic and hydrophobic antioxidants

A sensitive and simple procedure is described for evaluating the antioxidant activity of 21 known hydrophilic and hydrophobic compounds by using the chemiluminogenic reaction of lucigenin with hydrogen peroxide. The method was validated for linearity, limit of detection, precision, additivity and ruggedness and compared to the DPPH method. Furthermore, the IC(50) values of all compounds were calculated and compared by both methods and results are discussed on the basis of structure-antioxidant activity relationship. Among hydrophilic antioxidants, phenolic compounds with adjacent hydroxyl groups (gallic acid, caffeic acid, pyrocatechol, (+/-)-catechin, oleuropein) were found to be efficient antioxidants in chemiluminescence method with IC(50) values at the sub-micromolar range, while phenolic compounds without adjacent hydroxyl groups (p-coumaric acid, BHT, BHA, resorcinol) exhibited weak activity with IC(50) values>10microM. Alpha-tocopherol was the most efficient hydrophobic antioxidant (IC(50)=6.7microM) while oleic and linoleic acids were found to be very poor antioxidants (IC(50)>300microM). Finally the method was applied to the estimation of antioxidant activity of complex samples such as olive oils. To the best of our knowledge, this is the first time that total antioxidant activity can be determined directly in olive oil and not in aqueous extracts.

Flow-Based Methods with Chemiluminescence Detection for Food and Environmental Analysis: A Review

This paper presents an overview of flow-based methods in food and environmental analysis using chemiluminescence (CL) detection covering the period from 2005 to the present. The review discussses both automated flow methods of analysis [such as flow-injection analysis (FIA), sequential-injection analysis (SIA) and their variants] and separation techniques [liquid chromatography (LC) and capillary electrophoresis (CE) coupled to CL detection]. The most widely used CL reactions are presented together with representative applications in food and environmental analysis (determination of naturally occurring compounds, contaminants, additives as well as determination of inorganic and organic compounds).

Comparative Biophysical Studies of Sartan Class Drug Molecules Losartan and Candesartan (CV-11974) with Membrane Bilayers

The interactions of the antihypertensive AT(1) antagonists candesartan and losartan with membrane bilayers were studied through the application of DSC, Raman, and solid state (31)P NMR spectroscopies. (1)H and (13)C NMR resonances of candesartan were assigned on the basis of 1D and 2D NMR spectroscopy. A (31)P CP NMR broadline fitting methodology in combination with ab initio computations was implemented and, in conjunction with DSC and Raman results, provided valuable information regarding the perturbation, localization, orientation, and dynamic properties of the drugs in membrane models. In particular, results indicate that losartan anchors in the mesophase region of the lipid bilayers with the tetrazole group oriented toward the polar headgroup, whereas candesartan has less definite localization spanning from water interface toward the mesophase and upper segment of the hydrophobic region. Both sartan molecules decrease the mobilization of the phospholipids alkyl chains. Losartan exerts stronger interactions compared with candesartan, as depicted by the more prominent thermal, structural, and dipolar (1)H-(31)P changes that are caused in the lipid bilayers. At higher concentrations, candesartan strengthens the polar interactions and induces increased order at the bilayer surface. At the highest concentration used (20 mol %), only losartan induces formation of microdomains attributed to the flexibility of its alkyl chain. These results in correlation to reported data with other AT(1) antagonists strengthen the hypothesis that this class of molecules may approach the active site of the receptor by insertion in the lipid core, followed by lateral diffusion toward the binding site. Further, the similarities and differences of these drugs in their interactions with lipid bilayers establish, at least in part, their pharmacological properties.

Development of a generic assay for the determination of total trihydroxybenzoate derivatives based on gold-luminol chemiluminescence

A selective assay for the determination of one of the most important class of phenolic compounds, namely trihydroxybenzoates (monomeric and polymeric compounds having at least one gallate moiety) based on their enhancing effect on the chemiluminogenic reaction between gold ions and luminol is described for the first time. In the presence of trihydroxybenzoate derivatives, the light emission generated when alkaline luminol is oxidized by gold ions is amplified several orders of magnitude compared to other common phenolic compounds which exhibit minor reactivity or no reactivity at all (e.g. hydroxycinnamates, flavonols, benzenediols). Based on this property, the experimental conditions were optimized in order to enable the determination of total trihydroxybenzoates in complex mixtures without resorting to separation techniques. The method was applied to samples of different composition (teas, herbal infusions and wines) with satisfactory analytical features yielding detection limits at the 10(-7) mol L(-1) level, intra-day precision of 3.1%, inter-day precision less than 10% and recoveries between 88.7 and 97.6%. The strengths and weaknesses of the method were identified and discussed in relation to its application in real samples.

Luminescent Methods in the Analysis of Untreated Edible Oils: A Review

The present review describes the application of luminescent methods in the analysis of edible oils without any pretreatment such as extraction prior to analysis. Emphasis has been given to applications of chemiluminescence and fluorescence assays for determining quality parameters of edible oils, such as oxidative stability, antioxidant activity, and lipid hydroperoxides content, as well as classification or adulteration of vegetable oils.

Modified DPPH and ABTS Assays to Assess the Antioxidant Profile of Untreated Oils

This research dwells on two widely used spectrophotometric methods, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays, which assess the free radical scavenging activity (RSA) of natural samples and standard compounds. In particular, these assays were modified in order to simplify the evaluation of RSA of untreated edible oils, as well as to assess the antioxidant profile of oils’ hydrophilic and lipophilic extracts with the same analytical procedure. A thorough study highlighted the effect of solvents on the DPPH and ABTS methods and resulted in selecting 2-propanol and an ethanol/1-butanol solvent mixture as the reaction solvent for the DPPH method and the ABTS method, respectively. The developed methods were used to evaluate the RSA of 12 antioxidant compounds and 8 edible oils. Then, the contribution of lipophilic and hydrophilic extracts to the total RSA of oils was estimated. The obtained results demonstrate the applicability of the method to routine edible oil analysis.