Zulfiqur Ali

First human experiments with a novel non-invasive, non-optical continuous glucose monitoring system

This paper describes a non-invasive continuous glucose monitoring system based on impedance spectroscopy. Changes in the glucose concentrations can be monitored by varying the frequency in the radio band over a range, optimised to measure the impact of glucose on the impedance pattern. A number of clinical-experimental studies (hyperglycaemic excursions) were performed with healthy subjects in order to prove the applicability of this approach. The sensor used in these experiments is the size of a wristwatch and holds an open resonant circuit coupled to the skin and a circuit, performing an impedance measurement. In most cases, the experiments showed a good correlation between changes in blood glucose and the sensor recordings. A detailed description of the trials is presented. The results of this first series of experiments can be considered as a proof of concept for this novel non-invasive monitoring approach. Nevertheless, partly due to the indirect measurement, a considerable number of questions remain to be clarified.

Denuder Tubes for Sampling of Gaseous Species A Review

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Electrochemical immobilisation of enzymes. Part 4. - Co-immobilisation of glucose oxidase and ferro/ferricyanide in poly(N-methylpyrrole) films

Glucose oxidase and ferro/ferricyanide can be co-immobilised in poly(N-methylpyrrole) films. The two species compete for incorporation into the films so that on increasing the concentration of enzyme in the growth solution less ferro/ferricyanide is incorporated into the film, and vice versa. The ferro/ferricyanide entrapped within the film acts as a redox mediator for the oxidation of the enzyme at 0.45 V vs. SCE. Studies of the effect of variation of film thickness, glucose concentration, enzyme loading and ferro/ferricyanide loading are consistent with a model in which diffusion and enzyme-catalysed reaction within the film are coupled. Analysis of data from these experiments allows the rate parameters for these processes to be determined. These data are compared with previous results for oxygen mediation in poly(N-methylpyrrole) films.

Electrochemical detection of d-dimer as deep vein thrombosis marker using single-chain d-dimer antibody immobilized on functionalized polypyrrole

We describe a rapid and sensitive method for detection and quantification of d-dimer which is a biomarker present at elevated concentrations in patients with deep vein thrombosis (DVT) disorders. The method uses an immunosensor based on a single-chain antibody (ScAb) immobilized on a transducer surface and with a densely packed receptor layer. Detection is based on the redox activity of a N-alpha bis(carboxymethyl)-L-lysine (ANTA)/Cu2+ complex attached to a polypyrrole backbone. The resulting hybrid material: polypyrrole ANTA/metal complex/His-tag ScAb was characterized by AFM, surface plasmon resonance (SPR) and differential pulse voltammetry (DPV) for the optimization of the biosensor formation. The biosensor offers a promising template for antibody immobilization and for immunodetection of a specific D-dimer. The biosensor shows a remarkable variation in redox activity of the ANTA/Cu2+ complex after the D-dimer association with a binding constant Kd of 1 ng mL(-1). Electrochemical impedance spectroscopy (EIS) allows monitoring D-dimer association with a linear response between 0.1 ng mL(-1) and 500 ng mL(-1) and a detection limit of 100 pg mL(-1) in PBS is obtained. The biolayer exhibits the same sensitivity for the detection of d-dimer in human patient plasma samples. This assay method is versatile, offers enhanced performance for the evaluation of proteins association and could easily be extended to the detection of other proteins, present in serum human sample.

Application of the quartz crystal microbalance to the monitoring of Staphylococcus epidermidis antigen–antibody agglutination

The change in solution properties due to the agglutination of an antigen with its specific antibody has previously been used as a marker of infection. This method has been modified to allow the binding activity between species to be followed using the frequency response of a quartz crystal microbalance (QCM). The Bayston agglutination plate assay for Staphylococcus epidermidis has been modified to allow the electrode of a QCM to act as a direct sensor for the change in solution properties as agglutination occurs. Antibody and antigen were introduced to the crystal surface and the agglutination process was followed as a change in crystal resonant frequency. Serum, known to be infected with the organism, gave a titre of 3.9x10(-2)% v/v (-118 Hz, +/-12 SD, N = 9) matching that given by triplicate plate assay. Uninfected serum gave no frequency changes at this concentration, yielding a titre of 2.5x10(-2)% v/v again matching the plate titre (N = 3). Infected serum gave responses 40 times faster then those of the uninfected serum. The piezoelectric quartz crystal method gave a positive or negative diagnosis in <15 min compared with the 24 h required for the plate assay.

Production of rhamnolipid biosurfactants by Pseudomonas aeruginosa DS10-129 in a microfluidic bioreactor

A low‐cost μBR (microbioreactor) made from PTFE [poly(tetrafluoroethylene)] was used to cultivate a model organism, Pseudomonas aeruginosa DS10‐129. The progress of bioprocessing was monitored by comparing the growth of the organism in a μBR, a conventional bench scale bioreactor and a shake flask. Under the μBR conditions, the organism produced 23 mg/ml of pyocyanin that had antimicrobial effects against Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas teessidea and Pseudomonas clemancea. Furthermore, it produced a total of 106 μg/ml of effective biosurfactants consisting of dirhamnolipids (RL2) and monorhamnolipids (RL1). The biosurfactants reduced the surface tension of distilled water from 72 to 27.9 mN/m and emulsified kerosene by 71.30%. The pyocyanin and rhamnolipids were produced during the exponential and stationary phases of growth respectively. The results of the μBR were comparable to those obtained using conventional scale methods.

Broadband Cavity Enhanced Absorption Spectroscopy as a Detector for HPLC

The first demonstration of a cavity enhanced absorption spectroscopy (CEAS) based technique, applied to HPLC detection is reported. Broadband cavity enhanced absorption spectroscopy (BBCEAS) has been used for detection in a HPLC system (HPLC-BBCEAS). Measurements were made on the dyes rhodamine 6G and rhodamine B between 450 and 600 nm. The sensitivity of the measurements as determined by the minimum detectable change in the absorption coefficient, alpha(min), were 2.9 x 10(-5) cm(-1) at 527 nm and 1.9 x 10(-5) cm(-1) at 556 nm, the peak absorption wavelengths of rhodamine 6G and rhodamine B, respectively. The limits of detection (LOD) for the two dyes were 426 and 271 pM, respectively. The LOD of the HPLC-BBCEAS setup was found to be between 54 and 77 times lower than with a Perkin-Elmer HPLC (series 200) comprising a 200EP photodiode array detector. The sensitivity of the developed setup also compared favorably with the previous single wavelength HPLC-CRDS studies while using a considerably lower cost experimental setup and simpler experimental methodology. The use of BBCEAS detection also allowed the discrimination following an isocratic HPLC separation of the nearly co-eluting dyes rhodamine 6G and rhodamine B.

Liquid-phase broadband cavity-enhanced absorption spectroscopy measurements in a 2 mm cuvette.

A novel implementation of broadband cavity enhanced absorption spectroscopy (BBCEAS) has been used to perform sensitive visible wavelength measurements on liquid-phase solutions in a 2 mm cuvette placed at normal incidence to the cavity mirrors. The overall experimental methodology was simple, low cost, and similar to conventional ultraviolet-visible absorption spectroscopy. The cavity was formed by two concave high reflectivity mirrors. Three mirror sets with nominal reflectivities (R) of R = 0.99, 0.9945, and 0.999 were used. The light source consisted of a high intensity red, green, blue, or white LED. The detector was a compact charge-coupled device (CCD) spectrograph. Measurements were made on the representative analytes, Ho3+, and the dyes brilliant blue-R, sudan black, and coumarin 334 in appropriate solvents. Cavity enhancement factors (CEF) of up to 104 passes for the high reflectivity mirrors were obtained. The number of passes was limited by relatively high scattering and absorption losses in the cavity, of ∼1 × 10−2 per pass. Measurements over a wide wavelength range (420–670 nm) were also obtained in a single experiment with the white LED and the R = 0.99 mirror set for Ho3+ and sudan black. The sensitivity of the experimental setup could be determined by calculating the minimum detectable change in the absorption coefficient αmin. The values ranged from 5.1 × 10−5 to 1.2 × 10−3 cm−1. The limit of detection (LOD) for the strong absorber brilliant blue-R was 620 pM. A linear dynamic range of measurements of concentration over about two orders of magnitude was demonstrated. The overall sensitivity of the experimental setup compared very favorably with previous generally more experimentally complex and expensive liquid-phase cavity studies. Possible improvements to the technique and its applicability as an analytical tool are discussed.

Liquid-phase broadband cavity enhanced absorption spectroscopy (BBCEAS) studies in a 20 cm cell

Sensitive liquid-phase measurements have been made in a 20 cm cell using broadband cavity enhanced absorption spectroscopy (BBCEAS). The cavity was formed by two high reflectivity mirrors which were in direct contact with the liquid-phase analytes. Careful choice of solvent was required to minimise the effect of background solvent absorptions. Measurements were made on the broad absorber Sudan black, dissolved in acetonitrile, using a white LED light source and R > or = 0.99 cavity mirrors, leading to a cavity enhancement factor (CEF) of 82 at 584 nm. The sensitivity as measured by the minimum detectable change in the absorption coefficient (alpha(min)) was 3.4 x 10(-7) cm(-1). Further measurements were made on the strong absorber methylene blue dissolved in acetonitrile at 655 nm. A white LED was used with the R > or = 0.99 cavity mirrors, leading to a CEF of 78 and alpha(min) = 4.4 x 10(-7) cm(-1). The use of a more intense red LED also allowed measurements with higher reflectivity R > or = 0.999 cavity mirrors, leading to a CEF of 429 and alpha(min) = 2.8 x 10(-7) cm(-1). The sensitivity was limited by dark noise from the detector but nevertheless appears to represent the most sensitive liquid-phase absorption measurement to date.

Detection of bacterial contaminated milk by means of a quartz crystal microbalance based electronic nose

Headspace analysis by means of sensor arrays has been successfully applied to a wide range of qualitative applications. In this study, a six element array of coated Quartz Crystal Microbalance (QCM) sensors was used for the headspace analysis of milk volatiles. The sensors were exposed to uncontaminated samples of milk and samples contaminated with Pseudomonas fragi (Ps. fragi) or Escherichia coli (E. coli). Principal component analysis (PCA) was used to analyse the sensor array responses. No discrimination between uncontaminated milk samples and those contaminated with Ps. fragi was observed. This can be explained by Ps. fragi being a poor fermenter of milk. However, encouraging results were found for the discrimination between the milk samples and those contaminated with E. coli.