Real-time measurement of glucose using chrono-impedance technique on a second generation biosensor.

Chrono-impedance technique (CIT) was implemented as a new transduction method for real time measurement of glucose in a biosensor system based in carbon paste (CP)/Ferrocene (FC)/glucose oxidase (GOx). The system presents high selectivity because the optimal stimulation signal composed by a 165mV DC potential and 50mV(RMS) AC signal at 0.4Hz was used. The low DC potential used decreased the interfering species effect and the biosensor showed a linear impedance response toward glucose detection at concentrations from 0mM to 20mM,with 0.9853 and 0.9945 correlation coefficient for impedance module (|Z|) and phase (Φ), respectively. The results of quadruplicate sets reveal the high repeatability and reproducibility of the measurements with a relative standard deviation (RSD) less than 10%. CIT presented good accuracy (within 10% of the actual value) and precision did not exceed 15% of RSD for high concentration values and 20% for the low concentration ones. In addition, a high correlation coefficient (R(2)=0.9954) between chrono-impedance and colorimetric methods was obtained. On the other hand, when two samples prepared at the same conditions were measured in parallel with both methods (the measurement was repeated four times), it should be noticed that student's t-test produced no difference between the two mentioned methods (p=1). The biosensor system hereby presented is highly specific to glucose detection and shows a better linear range than the one reported on the previous article.

Evaluation of chrono-impedance technique as transduction method for a carbon paste/glucose oxidase (CP/GOx) based glucose biosensor.

The chrono-impedance technique (CIT) for real time determination of glucose concentration in a first generation glucose oxidase/carbon paste electrode was implemented. The biosensor was polarized with a signal composed of 900 mV DC potential and 50 mV(RMS) AC signal at 0.4 Hz. A frequency response analyzer was used to measure the complex impedance (magnitude |Z| and phase (Φ)) of the biosensor-bulk interface. Real time measurements were performed while glucose was added to the bulk within a concentration range of 0-40 mM. The cumulative impedance dose-response curves were used to construct calibration curves, both for magnitude and phase. The best fitting was obtained with a hyperbolic equation. Four biosensors were built obtaining five calibration curves for each of them. A single test measurement (unknown glucose concentration) was also obtained after each calibration procedure. Glucose concentrations were estimated with the calibration curves and also measured by colorimetry, the latter being the reference method. Besides, one-way ANOVA test evaluated repeatability. Difference between means was not statistically significant (p>0.01) for both magnitudes (|Z| and Φ). The Student's t-test assessed the differences significance, which produced in all cases p levels lower or equal than 0.44. Thus, CIT was proved to be a reliable method to measure glucose concentration in real time. Moreover, it showed high repeatability and compared well against colorimetry (r(2)=0.98).

Design and evaluation of a fast Fourier transform-based nonlinear dielectric spectrometer.

Nonlinear dielectric spectroscopy of micro-organism is carried out by applying a moderate electrical field to an aqueous sample through two metal electrodes. Several ad hoc nonlinear spectrometers were proposed in the literature. However, these designs barely compensated the nonlinear distortion derived from the electrode-electrolyte interfaces (EEI). Moreover, the contribution of the suspension is masked by the effect of the nonlinearity introduced by the electrode contacts. Conversely, the nonlinear capability of a commercial tetrapolar analyzer has not been fully investigated. In this paper a new nonlinear tetrapolar spectrometer is proposed based on a commercial linear apparatus and ad hoc control and signal processing software. The system was evaluated with discrete electronic phantoms and showed that it can measure nonlinear properties of aqueous suspension independently of the presence of EEI (ANOVA test, p>0.001). It was also tested with real aqueous samples. The harmonics observed in the current that circulates through the sample reveals useful information about the transfer function of the sample. The total harmonic distortion was computed for linear mediums. Values lower than -60 dB suggest that the system has enough capability to perform nonlinear microbiological analysis. Design specifications, sources of interference, and equipment's limitations are discussed.

Non-linear dielectric spectroscopy of microbiological suspensions.

BACKGROUND: Non-linear dielectric spectroscopy (NLDS) of microorganism was characterized by the generation of harmonics in the polarization current when a microorganism suspension was exposed to a sinusoidal electric field. The biological nonlinear response initially described was not well verified by other authors and the results were susceptible to ambiguous interpretation. In this paper NLDS was performed to yeast suspension in tripolar and tetrapolar configuration with a recently developed analyzer. METHODS: Tripolar analysis was carried out by applying sinusoidal voltages up to 1 V at the electrode interface. Tetrapolar analysis was carried on with sinusoidal field strengths from 0.1 V cm(-1) to 70 V cm(-1). Both analyses were performed within a frequency range from 1 Hz through 100 Hz. The harmonic amplitudes were Fourier-analyzed and expressed in dB. The third harmonic, as reported previously, was investigated. Statistical analysis (ANOVA) was used to test the effect of inhibitor an activator of the plasma membrane enzyme in the measured response. RESULTS: No significant non-linearities were observed in tetrapolar analysis, and no observable changes occurred when inhibitor and activator were added to the suspension. Statistical analysis confirmed these results.When a pure sinus voltage was applied to an electrode-yeast suspension interface, variations higher than 25 dB for the 3rd harmonic were observed. Variation higher than 20 dB in the 3rd harmonics has also been found when adding an inhibitor or activator of the membrane-bounded enzymes. These variations did not occur when the suspension was boiled. DISCUSSION: The lack of result in tetrapolar cells suggest that there is no, if any, harmonic generation in microbiological bulk suspension. The non-linear response observed was originated in the electrode-electrolyte interface. The frequency and voltage windows observed in previous tetrapolar analysis were repeated in the tripolar measurements, but maximum were not observed at the same values. CONCLUSION: Contrary to previous assertions, no repeatable dielectric non-linearity was exhibited in the bulk suspensions tested under the field and frequency condition reported with this recently designed analyzer. Indeed, interface related harmonics were observed and monitored during biochemical stimuli. The changes were coherent with the expected biological response.

Algorithm for identifying and separating beats from arterial pulse records.

BACKGROUND: This project was designed as an epidemiological aid-selecting tool for a small country health center with the general objective of screening out possible coronary patients. Peripheral artery function can be non-invasively evaluated by impedance plethysmography. Changes in these vessels appear as good predictors of future coronary behavior. Impedance plethysmography detects volume variations after simple occlusive maneuvers that may show indicative modifications in arterial/venous responses. Averaging of a series of pulses is needed and this, in turn, requires proper determination of the beginning and end of each beat. Thus, the objective here is to describe an algorithm to identify and separate out beats from a plethysmographic record. A secondary objective was to compare the output given by human operators against the algorithm. METHODS: The identification algorithm detected the beat's onset and end on the basis of the maximum rising phase, the choice of possible ventricular systolic starting points considering cardiac frequency, and the adjustment of some tolerance values to optimize the behavior. Out of 800 patients in the study, 40 occlusive records (supradiastolic- subsystolic) were randomly selected without any preliminary diagnosis. Radial impedance plethysmographic pulse and standard ECG were recorded digitizing and storing the data. Cardiac frequency was estimated with the Power Density Function and, thereafter, the signal was derived twice, followed by binarization of the first derivative and rectification of the second derivative. The product of the two latter results led to a weighing signal from which the cycles' onsets and ends were established. Weighed and frequency filters are needed along with the pre-establishment of their respective tolerances. Out of the 40 records, 30 seconds strands were randomly chosen to be analyzed by the algorithm and by two operators. Sensitivity and accuracy were calculated by means of the true/false and positive/negative criteria. Synchronization ability was measured through the coefficient of variation and the median value of correlation for each patient. These parameters were assessed by means of Friedman's ANOVA and Kendall Concordance test. RESULTS: Sensitivity was 97% and 91% for the two operators, respectively, while accuracy was cero for both of them. The synchronism variability analysis was significant (p < 0.01) for the two statistics, showing that the algorithm produced the best result. CONCLUSION: The proposed algorithm showed good performance as expressed by its high sensitivity. The correlation analysis demonstrated that, from the synchronism point of view, the algorithm performed the best detection. Patients with marked arrhythmic processes are not good candidates for this kind of analysis. At most, they would be singled out by the algorithm and, thereafter, to be checked by an operator.

Comparative analysis of hematocrit measurements by dielectric and impedance techniques.

In a previous paper, a new dielectric technique was used to estimate hematocrit (HTC) in extracorporeal blood circulation systems, independently of plasma conductivity or osmolarity. Although many impedance techniques have been formerly proposed in the literature, none has been evaluated against plasma conductivity and osmolarity. Herein, we estimate HTC based on permittivity changes and also with other four techniques found in the literature. Besides, the error incurred in each is also studied when plasma conductivity and osmolarity changed as much as 1 mS/cm and 50 mOsm/kg, respectively. The dielectric (permittivity) technique has an error close to 5.4%, while the others showed both tendencies, i.e., lower error (2.5%, two of them) and higher error (8.6% and 16.3%, the other two). The dielectric technique, even though did not produce the lowest error, provides a well-described physical model along with simple instrumentation.

Hematocrit measurement by dielectric spectroscopy.

Based on permittivity changes, a new method to measure hematocrit (HCT) in extracorporeal blood systems is presented. Human blood samples were tested at different HCT levels pairing the values of permittivity change, obtained by means of a commercial impedance analyzer, with traditional centrifugation measurements. Data were correlated using both linear and nonlinear regression. When using the lineal model, the comparison yielded a high correlation coefficient (r = 0.99). Theoretical simplifications suggest that the method is independent of changes in the conductivities of the intracellular and extracellular compartments. The influence of osmolarity and conductivity of the extracellular compartment was analyzed. It is shown that HCT can be predicted within an error lower than 5% when those parameters changed as much as 1 mS/cm and 50 mOsm/kg, respectively. Thus, the method appears as valid and viable showing good possibilities in applications such as renal dialysis.

Algoritmo de separación de latidos de pulso arterial / Separation algorithm arterial pulse beats

An algorithm is described to extract cardiac from a pletysmographic signal based on the maximum value of the second derivative. An initial set of points is detected and filtered with two consecutive temporal analyses...