Combined SPRi Sensor for Simultaneous Detection of Nitrate and Ammonium in Wastewater.

Water pollution is a serious problem in modern society. Agriculture, being responsible for the discharge of agrochemicals, organic matter, or drug residues, produces a huge amount of wastewater. Aquaponics has the potential to reduce both water consumption and the impact of water pollution on fish farming and plant production. In the aquatic environment, inorganic nitrogen is mostly present in the form of nitrate and ammonium ions. Nitrate, as a final product of ammonia mineralization, is the most common chemical contaminant in aquifers around the world. For continuous monitoring of nitrogen compounds in wastewater, we propose a sensor for the simultaneous detection of nitrate and ammonium. A surface plasmon resonance imaging method with enzyme-mediated detection was used. Active layers of nitrate reductase and glutamine synthetase were created on the gold surface of a biochip and tested for the sensing of nitrate and ammonium in water from an aquaponic system. The proposed sensor was applied in water samples with a concentration of NO3- and NH4+ in a range between 24-780 mg·L-1 and 0.26-120 mg·L-1, respectively, with minimal pretreatment of a sample by its dilution with a buffer prior to contact on a biochip surface.

A novel approach for measuring membrane permeability for organic compounds via surface plasmon resonance detection.

Well-known methods for measuring permeability of membranes include static or flow diffusion chambers. When studying the effects of organic compounds on plants, the use of such model systems allows to investigate xenobiotic behavior at the cuticular barrier level and obtain an understanding of the initial penetration processes of these substances into plant leaves. However, the use of diffusion chambers has disadvantages, including being time-consuming, requiring sampling, or a sufficiently large membrane area, which cannot be obtained from all types of plants. Therefore, we propose a new method based on surface plasmon resonance imaging (SPRi) to enable rapid membrane permeability evaluation. This study presents the methodology for measuring permeability of isolated cuticles for organic compounds via surface plasmon resonance detection, where the selected model analyte was the widely used pesticide metazachlor. Experiments were performed on the cuticles of Ficus elastica, Citrus pyriformis, and an artificial PES membrane, which is used in passive samplers for the detection of xenobiotics in water and soils. The average permeability for metazachlor was 5.23 × 10-14 m2 s-1 for C. pyriformis, 1.34 × 10-13 m2 s-1 for F. elastica, and 7.74 × 10-12 m2 s-1 for the PES membrane. We confirmed that the combination of a flow-through diffusion cell and real-time optical detection of transposed molecules represents a promising method for determining the permeability of membranes to xenobiotics occurring in the environment. This is necessary for determining a pesticide dosage in agriculture, selecting suitable membranes for passive samplers in analytics, testing membranes for water treatment, or studying material use of impregnated membranes.

Surface plasmon resonance: An innovative method for studying water permeability of plant cuticles.

The cuticle forms an effective barrier protecting plants from water loss. Its permeability to water and other compounds significantly differs between species, types of cuticle (stomatous, astomatous), and can be affected by a wide variety of ambient conditions. Enzymatic isolation of the leaf cuticle allows obtaining intact cuticles for permeability measurements. However, the most available gravimetric method, which is used for the assessment of water permeability of isolated cuticles, requires a relatively large area of the cuticle and does not allow the determination of membrane heterogeneity. We propose a new method for the determination of water permeance based on an on-line detection of water flux from a liquid phase to the atmosphere through isolated leaf cuticles in semi-flow chambers. This approach is new in using the phenomenon of surface plasmon resonance for the detection of the liquid phase refractive index affected by water vapor. Isolated cuticles of the leaves of Ficus elastica and an artificial polyethersulfone membrane were used for method evaluation. The composition of cuticular wax and its influence on cuticular permeability was also studied. It has been confirmed that the application of the surface plasmon resonance principle can be used for the assessment of leaf cuticle water permeability and heterogeneity.

A modified method for enzymatic isolation of and subsequent wax extraction from Arabidopsis thaliana leaf cuticle.

BACKGROUND: The plant cuticle represents one of the major adaptations of vascular plants to terrestrial life. Cuticular permeability and chemical composition differ among species. Arabidopsis thaliana is a widely used model for biochemical and molecular genetic studies in plants. However, attempts to isolate the intact cuticle from fresh leaves of Arabidopsis have failed so far. The goal of this study was to optimise an enzymatic method for cuticle isolation of species with a thin cuticle and to test it on several A. thaliana wild types and mutants. RESULTS: We developed a method for isolation of thin cuticles that allows reducing the isolation time, the separation of abaxial and adaxial cuticles, and avoids formation of wrinkles. Optical microscopy was used for studying cuticle intactness and scanning electron microscopy for visualisation of external and internal cuticle structures after isolation. Wax extracts were analysed by GC-MS. Isolation of intact cuticle was successful for all tested plants. The wax compositions (very-long-chained fatty acids, alcohols and alkanes) of intact leaves and isolated cuticles of wild type Col-0 were compared. CONCLUSIONS: We conclude that the optimised enzymatic method is suitable for the isolation of A. thaliana adaxial and abaxial cuticles. The isolated cuticles are suitable for microscopic observation. Analysis of wax composition revealed some discrepancies between isolated cuticles and intact leaves with a higher yield of wax in isolated cuticles.