A multichannel system integrating molecularly imprinted conductive polymers for ultrasensitive voltammetric determination of four steroid hormones in urine.

This work reports on a modularized electrochemical method for the determination of the hormones cortisol, progesterone, testosterone and 17ß-estradiol in urine. These hormones were employed as templates when generating molecular imprints from aniline and metanilic acid by electropolymerization on the surface of screen-printed electrodes. The electrically conductive imprint was characterized by SEM, AFM and cyclic voltammetry. A four-channel system was then established to enable simultaneous determination of the hormones by cyclic voltammetry. The detection limits for cortisol, progesterone, testosterone and 17ß-estradiol are as low as 2, 2.5, 10 and 9 ag·mL-1 (for S/N = 3). Graphical abstract A four-channel system was established to enable simultaneous determination of 4 steroid hormones by cyclic voltammetry and by using moleculalry imprinted polymers.

Novel 10-Bit Impedance-to-Digital Converter for Electrochemical Impedance Spectroscopy Measurements.

Electrochemical impedance spectroscopy (EIS) is a widely used technique in biomedical and chemical analysis. A novel 10-bit impedance-to-digital converter (IDC), which can measure and directly convert the magnitude and phase of impedance to digital codes, is proposed for the EIS measurement system. The proposed IDC is composed of a magnitude-to-digital converter (MDC) and a phase-to-digital converter (PDC). The proposed IDC was designed and fabricated using a 0.35 [Formula: see text] 2P4M mixed-signal polycide process, and the core area is only 0.07 mm2. Moreover, it can work over a very wide frequency range (0.1 mHz-100 kHz), and has excellent accuracy. According to the measured results, the DNL of the MDC is within -0.3/+0.3 LSB, and the INL is around -3/+1 LSB. Moreover, an EIS measurement system, which is composed of the proposed IDC chip and some other commercial chips, is built to measure ZoBell's and melatonin solutions for validating the function of the proposed IDC.

Electrochemical sensing of nuclear matrix protein 22 in urine with molecularly imprinted poly(ethylene-co-vinyl alcohol) coated zinc oxide nanorod arrays for clinical studies of bladder cancer diagnosis.

In 1996 and 2000, the US Food and Drug Administration (FDA) approved the use of Nuclear matrix protein 22 (NMP22) as a monitoring tool for predicting the recurrence/clearing of bladder cancer, and for screening undiagnosed individuals who have symptoms of, or are at risk for, that disease. The fabrication of electrodes for sensing NMP22 and their integration with a portable potentiostat in a homecare system may have great value. This work describes a sensing element comprised of molecularly imprinted polymers (MIPs) for the specific recognition of NMP22 target molecules. Zinc oxide (ZnO) nanorods (214 ± 45 nm in diameter and 1.08 ± 0.11 µm long) were hydrothermally grown on the sensing electrodes to increase the surface area to be coated with MIPs. A portable potentiostat was assembled and a data acquisition (DAQ) card and the Labview program were utilized to monitor electrochemical reaction to sense NMP22 in urine samples. Finally, in phase 0 clinical trials, measurements were made of samples from a few patients with bladder cancer using the NMP22 MIP-coated ZnO nanorods electrodes that were integrated into a portable potentiostat, revealing NMP 22 concentrations in the range 128 ± 19 to 588 ± 53 ng/mL.

Integrated potentiostat for electrochemical sensing of urinary 3-hydroxyanthranilic acid with molecularly imprinted poly(ethylene-co-vinyl alcohol).

Changing demographics, the rise of personalized medicine and increased identification of biomarkers for diagnosis and management of chronic disease have increased the demand for portable bioanalytical instrumentation and point-of-care. The recent development of molecularly imprinted polymers enables production of low cost and highly stable sensing chips; however, the commercially available and full functional instruments employed for electrochemical analysis have shortcomings in actual homecare applications. In this work, integrated circuits (ICs) for monolithic implementation of voltammeter potentiostat with a large dynamic current range (5 nA to 1.2 mA) and short conversion time (10 ms) were fabricated in a 0.35 µm complementary metal-oxide-semiconductor (CMOS) process. The new instrumentation was tested with molecular imprinted sensors for 3-hydroxyanthranilic acid (3HAA) in urine. The sensor consisted of molecular imprinted of poly(ethylene-co-vinyl alcohol)s (abbreviated as EVALs) for implementation in a flow injection analysis system. The EVAL containing 32 ethylene mol% had the highest imprinting effectiveness for the target molecules. Fit-for-purpose figures of merit were achieved with a limit-of-detection (LOD) of 3.06 pg/mL. The measurements obtained in real undiluted urine samples fell within the reference concentration range of 50-550 ng/mL.

Molecularly imprinted electrochemical sensing of urinary melatonin in a microfluidic system.

Melatonin levels may be related to the risks of breast cancer and prostate cancer. The measurement of urinary melatonin is also useful in monitoring serum melatonin levels following oral administration. In this work, melatonin is the target molecule, which is imprinted onto poly(ethylene-co-vinyl alcohol) by evaporation of the solvent on the working electrode of an electrochemical sensing chip. This sensing chip is used directly as a tool for optimizing the imprinting polymer composition, flow rate, and injection volume of the samples. Microfluidic sensing of the target and interference molecules revealed that the lowest detection limit is as low as ∼pM, and the electrochemical response is weak even at high interference concentrations. Poly(ethylene-co-vinyl alcohol), containing 44 mol. % ethylene, had an imprinting effectiveness of more than six-fold. In random urine analysis, the microfluidic amperometric measurements of melatonin levels with an additional and recovery of melatonin, the melatonin recovery achieved 94.78 ± 1.9% for melatonin at a concentration of 1.75-2.11 pg/mL.

Design of a dual-mode electrochemical measurement and analysis system.

A dual-mode electrochemical measurement and analysis system is proposed. This system includes a dual-mode chip, which was designed and fabricated by using TSMC 0.35 µm 3.3 V/5 V 2P4M mixed-signal CMOS process. Two electrochemical measurement and analysis methods, chronopotentiometry and voltammetry, can be performed by using the proposed chip and system. The proposed chip and system are verified successfully by performing voltammetry and chronopotentiometry on solutions.

Optical sensing of urinary melatonin with molecularly imprinted poly(ethylene-co-vinyl alcohol) coated zinc oxide nanorod arrays.

In this work, aligned zinc oxide (ZnO) nanorods were selectively hydrothermally grown on acetate-seeded spots on a gold substrate; the nanorods had an average length and diameter of 1.7µm and 240nm, respectively. Melatonin was imprinted into poly(ethylene-co-vinyl alcohol), EVAL, which was coated onto ZnO nanorod arrays. The ZnO nanorods not only increased the surface area for sensing target molecules, but also constituted an optical sensing element, as the ZnO fluorescence decreases when targets bind to the imprinted EVAL film; the fluorescence decrease, as a function of melatonin concentration, is well fit by a Langmuir adsorption isotherm. Poly(ethylene-co-vinyl alcohol) with 44mol% ethylene showed the best imprinting effectiveness (ratio of the fluorescence decrease on binding melatonin to imprinted vs. non-imprinted EVAL-coated ZnO nanorod arrays) among the several compositions studied. In real urine analysis, the MIP films responded linearly to added (exogenous) melatonin, even in the presence of many possible interfering compounds in urine. This demonstrates the feasibility of using these MIPs as part of a total urinalysis MIP system.

Sensing of digestive proteins in saliva with a molecularly imprinted poly(ethylene-co-vinyl alcohol) thin film coated quartz crystal microbalance sensor.

The quartz crystal microbalance (QCM) has a sensitivity comparable to that of the surface plasmon resonance (SPR) transducer. Molecularly imprinted polymers (MIPs) have a much lower cost than natural antibodies, they are easier to fabricate and more stable, and they exhibit satisfactory recognition ability when integrated onto sensing transducers. Hence, MIP-based QCM sensors have been used to recognize small molecules and, recently, microorganisms, but only a few have been adopted in protein sensing. In this work, a mixed salivary protein and poly(ethylene-co-vinyl alcohol), EVAL, solution is coated onto a QCM chip and a molecularly imprinted EVAL thin film formed by thermally induced phase separation (TIPS). The optimal ethylene mole ratios of the commercially available EVALs for the imprinting of amylase, lipase and lysozyme were found to be 32, 38, and 44 mol %, respectively. Finally, the salivary protein-imprinted EVAL-based QCM sensors were used to detect amylase, lipase and lysozyme in real samples (saliva) and their effectiveness was compared with that of a commercial ARCHITECT ci 8200 chemical analysis system. The limits of detection (LOD) for those salivary proteins were as low as ∼pM.

A portable electrochemical sensor for caffeine and (-)epigallocatechin gallate based on molecularly imprinted poly(ethylene-co-vinyl alcohol) recognition element.

The U.S. Food and Drug Administration allows a maximum of 72 mg of caffeine per 12 oz. serving (6 mg/oz). Consuming 400 mg of caffeine 3 times daily for 7 days may develop sleep disruption effects. However, it is still very hard for people to estimate how much caffeine is intake daily. Moreover, (-)epigallocatechin gallate (EGCG) is studied a potent antioxidant that may have therapeutic properties for anti-aging and cancer. Conventionally, both caffeine and EGCG could be measured by the protocols of high performance liquid chromatography; however, high precision instruments are required. In this work, the caffeine and EGCG are used as the template molecules and imprinted into poly(ethylene-co-vinyl alcohol), EVAL, via solvent evaporation. The EVAL membrane is then used as the sensing element for electrochemical analysis after templates removal. From the cyclic voltammetry measurement of the caffeine, the peak oxidation potential is shifted from 0.36 to 0.46 V when the final concentration of caffeine is from 0.01 to 1 mg/mL, and the highest current density is about 0.18 microA/cm2. The caffeine and EGCG concentrations measured in three real samples are about 0.10-0.13 mg/mL and 0.49-1.74 mg/mL, respectively. This molecularly imprinted polymeric coated electrode is potential employed as a home-care system.

Formation and recognition characteristics of albumin-imprinted poly(ethylene-co-vinyl-alcohol) membranes.

The recognition of proteins by molecular imprinted polymers (MIPs) is an area of great current interest. Protein-imprinted polymers, or artificial antibodies, with high recognition capability could be used as the sensing material in a protein microarray, or for protein separation for clinical or research applications. However, a great concern in the synthesis of protein MIPs is often the cost of template protein. Surface-based molecular imprinting could offer the advantage of lower usage of template protein, as well overcoming problematic protein solubility in monomers. In addition, the microcontact imprinting method may increase the reproducibility of MIP film formation. In this study, a novel method to prepare the protein imprinted polymers was demonstrated using the phase inversion formation of poly(ethylene-co-vinyl-alcohol), EVAL, membranes with the model protein albumin. EVAL polymer solution (25 wt% of EVAL in dimethyl sulfoxide, DMSO) was cast on glass slides with immobilized template proteins, and MIPs were then solidified by evaporation of DMSO. Several factors were shown to affect the recognition ability of the MIPs, including the template concentration, binding temperature and with the presence of other proteins and biomolecules in the samples.

Breast tumor classification of ultrasound images using a reversible round-off nonrecursive 1-D discrete periodic wavelet transform.

The infiltrative nature of lesions is a significant feature of malignant breast lesion on ultrasound image. Characterizing the infiltrative nature of lesions with computationally inexpensive and highly efficacious features is crucial for the realization of a computer-aided diagnosis system. In this study, the infiltrative nature is regarded as an energy that produces irregularly and considerably local variances in a 1-D signal. The local variances can be characterized by a few high octave energies (i.e., the channel energies close to low-frequency bands) in a 1-D discrete periodic wavelet transform. For computational cost reduction, high octave decomposition is performed by a reversible round-off 1-D nonrecursive discrete periodic wavelet transform. A test dataset of breast sonograms with the lesion contour delineated by an experienced physician and two inexperienced persons is built for feature efficacy evaluation. High individual performance results imply that the proposed feature is well correlated with the diagnosis of the experienced physician. Experimental results also reveal that with a great performance improvement, the proposed feature is suitable for the combination with some morphometric parameters.

Urinalysis with molecularly imprinted poly(ethylene-co-vinyl alcohol) potentiostat sensors.

Among many important biomarkers excreted in urine are albumin, uric acid, glucose, urea, creatine and creatinine. In the growing elderly population, these biomarkers may be useful correlates with kidney dysfunction, infection and related problems such as glomerular, proximal, and distal convoluted tubule functions, diabetes, hypertension and proteinuria. This study employed solvent evaporation processing of poly(ethylene-co-vinyl alcohol), (EVAL) to form molecularly imprinted polymers (MIPs) that recognize creatinine, urea, and lysozyme. The mole ratio of ethylene to vinyl alcohol affected the performance: 27 mol% ethylene gave the highest imprinting effectiveness for creatinine and urea, while 44 mol% gave the highest effectiveness for lysozyme. Electrochemical examination using a home made potentiostat and imprinted polymer electrode showed electrical signals responsive to the target molecules. Finally, an actual urine sample was tested using the electrode. The test results were compared with those of the commercial instrument ARCHITECT ci 8200 system to precisely determine the accuracy of the molecularly imprinted polymer electrode for urinalysis.

A portable potentiostat for the bilirubin-specific sensor prepared from molecular imprinting.

A portable amperometric potentiostat was designed and implemented in this work. It was developed to acquisit the current signals produced from bilirubin by an electrochemical sensor. Based on an SOC-based chip, this potentiostat has the merits of moderate accuracy, small size, low cost, and high portability. The bilirubin electrode was prepared by synthesizing a thin layer of bilirubin imprinted poly(methacrylic acid-co-ethylene glycol dimethacrylate) onto the Au layer. With the molecularly imprinted polymer (MIP) film, specific detection of bilirubin was successfully achieved. The cyclic voltammogram of the electrode was measured from this assembled potentiostat. The performance from a commercial potentiostat was considered rather stable and was used as a reference to examine and evaluate the performance of the assembled potentiostat. The detected current signals by the bilirubin sensing were obtained. Linear calibration with a sensitivity of 1.344+/-0.38 microA/mg dl was achieved. Our experimental results showed that the proposed potentiostat's performance could achieve sufficient performance. The evaluation was also made from the aspects such as reset time and steady-response time. The self-assembled potentiostat thus demonstrated its ability in precise detection of bilirubin from an electrode layered with the imprinted polymer film.

A low-voltage wide-input CMOS comparator for sensor application using back-gate technique.

In this paper, two new analog CMOS comparators (Type-I and Type-II) with low-voltage and wide-input capabilities are proposed. The comparator receives two analog inputs and puts out one digital state to identify the larger (or the smaller) of the input variables, which represents an useful operation in data conversion and sensory signal processing. Without using special fabrication technologies, the supply voltage of the circuit is reduced to 1 V. Due to the utilization of CMOS back-gate technique, the input range of the comparators is greatly improved. The comparators are composed of bulk-driven stage and dynamic latch. By using a CMOS n-well technology, the results of HSPICE simulations indicate that the response time of Type-I circuit is 1 micros under 10 mV identified resolution. Type-II comparator achieves 5 mV identified resolution. The input dynamic ranges of the comparators are approximately rail-to-rail.