AirQuality Lab-on-a-Drone: A Low-Cost 3D-Printed Analytical IoT Platform for Vertical Monitoring of Gaseous H2S.

The measurement of gaseous compounds in the atmosphere is a multichallenging task due to their low concentration range, long and latitudinal concentration variations, and the presence of sample interferents. Herein, we present a quadcopter drone deployed with a fully integrated 3D-printed analytical laboratory for H2S monitoring. Also, the analytical system makes part of the Internet of Things approach. The analytical method applied was based on the reaction between fluorescein mercuric acetate and H2S that led to fluorescence quenching. A 5 V micropump at a constant airflow of 50 mL min-1 was employed to deliver constant air into a flask containing 800 µL of the reagent. The analytical signal was obtained using a light-emitting diode and a miniaturized digital light detector. The method enabled the detection of H2S in the range from 15 to 200 ppbv, with a reproducibility of 5% for a sampling time of 10 min and an limit of detection of 9 ppbv. All devices were controlled using an Arduino powered by a small power bank, and the results were transmitted to a smartphone via Bluetooth. The proposed device resulted in a weight of 300 g and an overall cost of ∼50 USD. The platform was used to monitor the concentration of H2S in different intervals next to a wastewater treatment plant at ground and vertical levels. The ability to perform all analytical steps in the same device, the low-energy requirements, the low weight, and the attachment of data transmission modules offer new possibilities for drone-based analytical systems for air pollution monitoring.

Nitrite Determination in Environmental Water Samples Using Microchip Electrophoresis Coupled with Amperometric Detection.

Nitrite is considered an important target analyte for environmental monitoring. In water resources, nitrite is the result of the nitrogen cycle and the leaching processes of pesticides based on nitrogenous compounds. A high concentration of nitrite can be associated with intoxication processes and metabolic disorders in humans. The present study describes the development of a portable analytical methodology based on microchip electrophoresis coupled with amperometric detection for the determination of nitrite in environmental water samples. Electrophoretic and detection conditions were optimized, and the best separations were achieved within 60 s by employing a mixture of 30 mmol L-1 lactic acid and 15 mmol L-1 histidine (pH = 3.8) as a running buffer applying 0.7 V to the working electrode (versus Pt) for amperometric measurements. The developed methodology revealed a satisfactory linear behavior in the concentration range between 20 and 80 µmolL-1 (R2 = 0.999) with a limit of detection of 1.3 µmolL-1. The nitrite concentration was determined in five water samples and the achieved values ranged from (28.7 ± 1.6) to (67.1 ± 0.5) µmol L-1. The data showed that using the proposed methodology revealed satisfactory recovery values (83.5-103.8%) and is in good agreement with the reference technique. Due to its low sample consumption, portability potential, high analytical frequency, and instrumental simplicity, the developed methodology may be considered a promising strategy to monitor and quantitatively determine nitrite in environmental samples.

Molecularly imprinted polymer as sorbent phase for disposable pipette extraction: A potential approach for creatinine analysis in human urine samples.

Traditionally, creatinine determination is made by a spectrophotometric method; however, some compounds present in biological samples can interfere with creatinine determination, decreasing the sensitivity of the method in urine samples. Consequently, we report the development of a new molecularly imprinted polymer as a sorbent phase for disposable pipette extraction to determine creatinine in urine samples by high-performance liquid chromatography with UV detection. The synthesized polymer showed a high superficial area and presented a first-order kinetic reaction and a high selectivity for creatinine extraction compared to the non-molecularly imprinted polymer. The main disposable pipette extraction variables evaluated included the number of draw/eject cycles, the pH of the solution and desorption solvent type. The developed method showed an inter and intra-day precision from 1.3% to 2.0% and 0.8-1.6% respectively, accuracy values ranging from 82.3% to 102.1% respectively and recovery values ranging between 96.5% and 101.3%, with a limit of quantification of 0.302 g L-1. The application of the developed method in real urine samples showed creatinine concentrations ranging from 0.55 to 6.61 g L-1. Thus, the developed method was revealed to be an efficient strategy for creatinine determination, reducing analysis time (3 min) and solvent use, and increasing selectivity compared with DPX commercial sorbents.

Organic beet leaves and stalk juice attenuates HDL-C reduction induced by high-fat meal in dyslipidemic patients: A pilot randomized controlled trial.

OBJECTIVES: Beet leaves and stalks are rich in polyphenols; however, their effect on risk factors for cardiovascular disease in humans, to our knowledge, has not yet been investigated. The aim of this study was to analyze the acute effect of beet leaves and stalk juice, containing different concentrations of polyphenols, on lipemia, glycemic control, nitric oxide concentration, and blood pressure in patients with dyslipidemia after a high-fat meal. METHODS: In a randomized double-blind, placebo-controlled, crossover pilot study, patients 20 to 59 y of age with dyslipidemia were fed a single high-fat meal supplemented with either a placebo or one of two organic beet leaves and stalk juices rich in polyphenols (32 or 77.5 mg EAG/100 mL) with a 1-wk washout. Thus, each group was composed of 13 patients. Blood samples were obtained at fasting and 30, 60, 120, and 180 min after intervention. Total cholesterol, high-density lipoprotein, triacylglycerols, glucose, insulin, nitrite and nitrate, and blood pressure were assessed at each time period. The high-fat meal increased triacylglycerol levels after 120 (P < 0.001) and 180 min (P < 0.001) and reduced high-density lipoprotein cholesterol after 60 min (P < 0.05). This reduction was attenuated in both groups that received BLS juices after 120 min (P = 0.005). A reduction in diastolic blood pressure within groups that received BLS juice was also observed. RESULTS: There was no significant difference between groups for other biomarkers. CONCLUSION: The beet leaves and stalk juice attenuated the reduction of high-density lipoprotein cholesterol induced by a high-fat meal.

Role of the Carotid Bodies in the Hypertensive and Natriuretic Responses to NaCl Load in Conscious Rats.

Hyperosmotic challenges trigger a hypertensive response and natriuresis mediated by central and peripheral sensors. Here, we evaluated the importance of the carotid bodies for the hypertensive and natriuretic responses to acute and sub-chronic NaCl load in conscious rats. Male Wistar rats (250-330 g) submitted to bilateral carotid body removal (CBX) or sham surgery were used. One day after the surgery, the changes in arterial blood pressure (n = 6-7/group) and renal sodium excretion (n = 10/group) to intravenous infusion of 3 M NaCl (1.8 mL/kg b.w. during 1 min) were evaluated in non-anesthetized rats. Another cohort of sham (n = 8) and CBX rats (n = 6) had access to 0.3 M NaCl as the only source of fluid to drink for 7 days while ingestion and renal excretion were monitored daily. The sodium balance was calculated as the difference between sodium infused/ingested and excreted. CBX reduced the hypertensive (8 ± 2 mmHg, vs. sham rats: 19 ± 2 mmHg; p < 0.05) and natriuretic responses (1.33 ± 0.13 mmol/90 min, vs. sham: 1.81 ± 0.11 mmol/90 min; p < 0.05) to acute intravenous infusion of 3 M NaCl, leading to an increase of sodium balance (0.38 ± 0.11 mmol/90 min, vs. sham: -0.06 ± 0.10 mmol/90 min; p < 0.05). In CBX rats, sub-chronic NaCl load with 0.3 M NaCl to drink for 7 days increased sodium balance (18.13 ± 4.45 mmol, vs. sham: 5.58 ± 1.71 mmol; p < 0.05) and plasma sodium concentration (164 ± 5 mmol/L, vs. sham: 140 ± 7 mmol/L; p < 0.05), without changing arterial pressure (121 ± 9 mmHg, vs. sham: 116 ± 2 mmHg). These results suggest that carotid bodies are important for the maintenance of the hypertensive response to acute hypertonic challenges and for sodium excretion to both acute and chronic NaCl load.

Microfluidic toner-based analytical devices: disposable, lightweight, and portable platforms for point-of-care diagnostics with colorimetric detection.

This chapter describes the development of microfluidic toner-based analytical devices (µTADs) to perform clinical diagnostics using a scanner or cell-phone camera. µTADs have been produced in a platform composed of polyester and toner by the direct-printing technology (DPT) in a matter of minutes. This technology offers simplicity and versatility, and it does not require any sophisticated instrumentation. Toner-based devices integrate the current generation of disposable analytical devices along paper-based chips. The cost of one µTAD has been estimated to be lower than $0.10. In addition, these platforms are lightweight and portable thus enabling their use for point-of-care applications. In the last 5 years, great efforts have been dedicated to spread out the use of µTADs in bioassays. The current chapter reports the fabrication of printed microplates and integrated microfluidic toner-based devices for dengue diagnostics and rapid colorimetric assays with clinically relevant analytes including cholesterol, triglycerides, total proteins, and glucose. The use of µTADs associated with cell-phone camera may contribute to the health care, in special, to people housed in developing regions or with limited access to clinics and hospitals.

Rational selection of substrates to improve color intensity and uniformity on microfluidic paper-based analytical devices.

A systematic investigation was conducted to study the effect of paper type on the analytical performance of a series of microfluidic paper-based analytical devices (µPADs) fabricated using a CO2 laser engraver. Samples included three different grades of Whatman chromatography paper, and three grades of Whatman filter paper. According to the data collected and the characterization performed, different papers offer a wide range of flow rate, thickness, and pore size. After optimizing the channel widths on the µPAD, the focus of this study was directed towards the color intensity and color uniformity formed during a colorimetric enzymatic reaction. According to the results herein described, the type of paper and the volume of reagents dispensed in each detection zone can determine the color intensity and uniformity. Therefore, the objective of this communication is to provide rational guidelines for the selection of paper substrates for the fabrication of µPADs.

Laser-printing of toner-based 96-microzone plates for immunoassays.

This work describes the quick and simple fabrication of toner-based 96-microzone plates by a direct-printing technology. The printer deposits a toner layer (ca. 5 µm thick) on the polyester surface which acts as a hydrophobic barrier to confine small volumes of sample on test zones (wells). A 96-microzone toner plate was explored to demonstrate its capability of performing enzyme-linked immunosorbent assay (ELISA). The detection of anti-immunoglobulin G (anti-IgG) and immunoglobulin M (IgM) antibodies has been successfully achieved in cell culture and serum samples, respectively. The use of a conventional microplate reader has allowed obtaining a limit of detection of 13 fmol of mouse IgG per zone on printed microplates. The IgM antibody has been detected in a serum sample collected from a patient infected with dengue virus. The detection of a primary infection has been provided by a microplate reader and also by a cell phone camera. Besides the bioanalytical feasibility, toner-based zones have shown good repeatability for inter-zone and intra-plate comparisons. The relative standard deviation (RSD) values for inter-zone (n = 12) and intra-plate (n = 3) comparisons were lower than 6% and 11%, respectively. Furthermore, it was found that the lifetime of each printed microplate depends on the storage temperature. The shelf life for devices stored at 10 °C has been estimated to be ca. four weeks.

Capillary-driven toner-based microfluidic devices for clinical diagnostics with colorimetric detection.

The fabrication of toner-based microfluidic devices to perform clinical diagnostics with capillary action and colorimetric detection is described in this report. Test zones and microfluidic channels were drawn in a graphic software package and laser printed on a polyester film. The printed layout and its mirror image were aligned with an intermediary cut-through polyester film and then thermally laminated together at 150 °C at 60 cm/min to obtain a channel with ca. 100-µm depth. Colorimetric assays for glucose, protein, and cholesterol were successfully performed using a desktop scanner. The limit of detection (LD) values found for protein, cholesterol, and glucose were 8, 0.2, and 0.3 mg/mL, respectively. The relative standard deviation (RSD) values for an interdevices comparison were 6%, 1%, and 3% for protein, cholesterol, and glucose, respectively. Bioassays were successfully performed on toner-based devices stored at different temperatures during five consecutive days without loss of activity.

High-voltage power supplies to capillary and microchip electrophoresis.

Over the past years, the development of capillary electrophoresis (CE) and microchip electrophoresis (ME) systems has grown due to instrumental simplicity and wide application. In both CE and ME, the application of a high voltage (HV) is a crucial step in the electrokinetic (EK) injection and separation processes. Particularly on ME devices, EK injection is often performed with three different modes: gated, pinched, and unpinched. In all these cases, different potential values may be applied to one or multiple channels to control the injection of small sample volumes as well as the separation process. For this reason, the construction of reliable HV power supplies (HVPS) is required. This review covers the advances of the development of commercial and laboratory-built HVPS for CE and ME. Moreover, it intends to be a guide for new developers of electrophoresis instrumentation.

Doping of a dielectric layer as a new alternative for increasing sensitivity of the contactless conductivity detection in microchips.

This communication describes a new procedure to increase the sensitivity of C(4)D in PDMS/glass microchips. The method consists in doping the insulating layer (PDMS) over the electrodes with nanoparticles of TiO(2), increasing thus its dielectric constant. The experimental protocol is simple, inexpensive, and fast.

Toner and paper-based fabrication techniques for microfluidic applications.

The interest in low-cost microfluidic platforms as well as emerging microfabrication techniques has increased considerably over the last years. Toner- and paper-based techniques have appeared as two of the most promising platforms for the production of disposable devices for on-chip applications. This review focuses on recent advances in the fabrication techniques and in the analytical/bioanalytical applications of toner and paper-based devices. The discussion is divided in two parts dealing with (i) toner and (ii) paper devices. Examples of miniaturized devices fabricated by using direct-printing or toner transfer masking in polyester-toner, glass, PDMS as well as conductive platforms as recordable compact disks and printed circuit board are presented. The construction and the use of paper-based devices for off-site diagnosis and bioassays are also described to cover this emerging platform for low-cost diagnostics.

A rapid and reliable bonding process for microchip electrophoresis fabricated in glass substrates.

In this report, we describe a rapid and reliable process to bond channels fabricated in glass substrates. Glass channels were fabricated by photolithography and wet chemical etching. The resulting channels were bonded against another glass plate containing a 50-microm thick PDMS layer. This same PDMS layer was also used to provide the electrical insulation of planar electrodes to carry out capacitively coupled contactless conductivity detection. The analytical performance of the proposed device was shown by using both LIF and capacitively coupled contactless conductivity detection systems. Efficiency around 47,000 plates/m was achieved with good chip-to-chip repeatability and satisfactory long-term stability of EOF. The RSD for the EOF measured in three different devices was ca. 7%. For a chip-to-chip comparison, the RSD values for migration time, electrophoretic current and peak area were below 10%. With the proposed approach, a single chip can be fabricated in less than 30 min including patterning, etching and sealing steps. This fabrication process is faster and easier than the thermal bonding process. Besides, the proposed method does not require high temperatures and provides excellent day-to-day and device-to-device repeatability.

Comparison of the analytical performance of electrophoresis microchannels fabricated in PDMS, glass, and polyester-toner.

This paper compares the analytical performance of microchannels fabricated in PDMS, glass, and polyester-toner for electrophoretic separations. Glass and PDMS chips were fabricated using well-established photolithographic and replica-molding procedures, respectively. PDMS channels were sealed against three different types of materials: native PDMS, plasma-oxidized PDMS, and glass. Polyester-toner chips were micromachined by a direct-printing process using an office laser printer. All microchannels were fabricated with similar dimensions according to the limitations of the direct-printing process (width/depth 150 microm/12 microm). LIF was employed for detection to rule out any losses in separation efficiency due to the detector configuration. Two fluorescent dyes, coumarin and fluorescein, were used as model analytes. Devices were evaluated for the following parameters related to electrophoretic separations: EOF, heat dissipation, injection reproducibility, separation efficiency, and adsorption to channel wall.

Fabrication and integration of planar electrodes for contactless conductivity detection on polyester-toner electrophoresis microchips.

In this report, we describe the microfabrication and integration of planar electrodes for contactless conductivity detection on polyester-toner (PT) electrophoresis microchips using toner masks. Planar electrodes were fabricated by three simple steps: (i) drawing and laser-printing the electrode geometry on polyester films, (ii) sputtering deposition onto substrates, and (iii) removal of toner layer by a lift-off process. The polyester film with anchored electrodes was integrated to PT electrophoresis microchannels by lamination at 120 degrees C in less than 1 min. The electrodes were designed in an antiparallel configuration with 750 microm width and 750 microm gap between them. The best results were recorded with a frequency of 400 kHz and 10 Vpp using a sinusoidal wave. The analytical performance of the proposed microchip was evaluated by electrophoretic separation of potassium, sodium and lithium in 150 microm wide x 6 microm deep microchannels. Under an electric field of 250 V/cm the analytes were successfully separated in less than 90 s with efficiencies ranging from 7000 to 13,000 plates. The detection limits (S/N = 3) found for K+, Na+, and Li+ were 3.1, 4.3, and 7.2 micromol/L, respectively. Besides the low-cost and instrumental simplicity, the integrated PT chip eliminates the problem of manual alignment and gluing of the electrodes, permitting more robustness and better reproducibility, therefore, more suitable for mass production of electrophoresis microchips.

Polyurethane from biosource as a new material for fabrication of microfluidic devices by rapid prototyping.

This paper presents the use of elastomeric polyurethane (PU), derived from castor oil (CO) biosource, as a new material for fabrication of microfluidic devices by rapid prototyping. Including the irreversible sealing step, PU microchips were fabricated in less than 1h by casting PU resin directly on the positive high-relief molds fabricated by standard photolithography and nickel electrodeposition. Physical characterization of microchannels was performed by scanning electron microscopy (SEM) and profilometry. Polymer surface was characterized using contact angle measurements and the results showed that the hydrophilicity of the PU surface increases after oxygen plasma treatment. The polymer surface demonstrated the capability of generating an electroosmotic flow (EOF) of 2.6 x 10(-4)cm(2)V(-1)s(-1) at pH 7 in the cathode direction, which was characterized by current monitoring method at different pH values. The compatibility of PU with a wide range of solvents and electrolytes was tested by determining its degree of swelling over a 24h period of contact. The performance of microfluidic systems fabricated using this new material was evaluated by fabricating miniaturized capillary electrophoresis systems. Epinephrine and l-DOPA, as model analytes, were separated in aqueous solutions and detected with end-channel amperometric detection.

A toner-mediated lithographic technology for rapid prototyping of glass microchannels.

A simple, fast, and inexpensive masking technology without any photolithographic step to produce glass microchannels is proposed in this work. This innovative process is based on the use of toner layers as mask for wet chemical etching. The layouts were projected in graphic software and printed on wax paper using a laser printer. The toner layer was thermally transferred from the paper to cleaned glass surfaces (microscope slides) at 130 degrees C for 2 min. After thermal transference, the glass channel was etched using 25% (v/v) hydrofluoric acid (HF) solution. The toner mask was then removed by cotton soaked in acetonitrile. The etching rate was approximately 7.1 +/- 0.6 microm min(-1). This process is economically more attractive than conventional methods because it does not require any sophisticated instrumentation and it can be implemented in any chemical/biochemical laboratory. The glass channel was thermally bonded against a flat glass cover and its analytical feasibility was investigated using capacitively coupled contactless conductivity detection (C(4)D) and laser-induced fluorescence (LIF) detection.

Electrokinetic control of fluid in plastified laser-printed poly(ethylene terephthalate)-toner microchips.

The application of plastified laser-printed poly(ethylene terephthalate)(PET)-toner microchips to capillary electrophoresis was investigated. Electroosmotic flow was observed in the direction of the cathode for the buffer system studied (phosphate, pH 3-10). Average electroosmotic mobilities of 1.71 x 10(-4) to 4.35 x 10(-4) cm(2) V(-1) s(-1) were observed from pH 3 to 10. This variation suggests that silica fillers in the toner and on the surface of the polymer dominate the zeta potential of the material, which is also confirmed by XPS measurements. Dopamine and catechol were used as model analytes for microchip electrophoresis in combination with electrochemical detection. Results show that these two analytes can be efficiently separated and detected electrochemically with the plastified laser-printed PET-toner microchips.