Development of a gas-diffusion microfluidic paper-based analytical device (µPAD) for the determination of ammonia in wastewater samples.

An inexpensive, disposable and highly selective microfluidic paper-based analytical device (µPAD) is described for the determination of ammonia (molecular ammonia and ammonium cation) in wastewaters which implements for the first time a gas-diffusion separation step on a paper-based platform. Its hydrophilic reagent zones were defined by printing filter paper with a hydrophobic paper sizing agent using a conventional inkjet printer. The sample was introduced into the sodium hydroxide impregnated sample zone of the µPAD. This allowed the quantitative conversion of the ammonium ion to molecular ammonia which diffused across the hydrophobic microporous Teflon membrane of the device into an adjacent hydrophilic reagent zone containing the acid-base indicator 3-nitrophenol or bromothymol blue. The change in indicator color was measured using a desktop scanner for ammonia quantification. Under optimal conditions, the µPAD is characterized by a limit of detection of 0.8 and 1.8 mg N L(-1) and repeatability of 3.1 and 3.7% (n ≥ 10, 20 mg N L(-1)), expressed as relative standard deviation, in the case of 3-nitrophenol or bromothymol blue, respectively. This µPAD was used successfully for the determination of ammonia in sewage and soil water samples. The small dimensions, minimal reagent consumption, low cost, simplicity of operation, and possibility of using a portable scanner make the proposed µPAD suitable for on-site ammonia monitoring in contaminated environmental waters and domestic, agricultural and industrial wastewaters. The successful implementation of the gas-diffusion approach on a paper-based platform is expected to result in the development of other µPADs for volatile analytes.

Microfluidic paper-based analytical device for the determination of nitrite and nitrate.

A low-cost disposable colorimetric microfluidic paper-based analytical device (µPAD) was developed for the determination of nitrite and nitrate. Nitrite is determined directly by the Griess reaction while nitrate is first reduced to nitrite in a hydrophilic channel of the µPAD with immobilized zinc microparticles. This µPAD is fabricated by a simple and inexpensive inkjet printing method. Under optimal conditions, the limits of detection and quantification for nitrite are 1.0 and 7.8 µM, respectively, while the corresponding values for nitrate are 19 and 48 µM, respectively. The repeatability, expressed as relative standard deviation (RSD), is less than 2.9% and 5.6% (n ≤ 8) for the determination of nitrite and nitrate, respectively. This µPAD was successfully applied to the determination of nitrate and nitrite in both synthetic and natural water samples. It is user and environmentally friendly and suitable for on-site measurement of the analytes mentioned above in environmental and drinking waters.

Microfluidic paper-based analytical device for the speciation of inorganic nitrogen species.

A microfluidic paper-based analytical device (µPAD) utilizing gas-diffusion separation and solid-phase reduction was developed for the first time for the determination of both ammonium and nitrate, which are the dominant inorganic nitrogen species in environmental waters. The device consists of 3 filter paper layers accommodating the sample, reagent and detection zones. The reagent zone is separated from the detection zone by a semipermeable hydrophobic membrane and acts as a solid-phase reactor where nitrate is reduced to ammonia by Devarda's alloy microparticles, integrated into a µPAD for the first time. The detection zone incorporates the acid-base indicators bromothymol blue (BTB) or nitrazine yellow (NY) and changes colour in two steps. Initially the colour change is caused by ammonia generated by the reaction of ammonium and sodium hydroxide in the sample zone. This colour change is followed by a subsequent colour change as a result of the ammonia produced by the reduction of nitrate by the Devarda's alloy microparticles. The corresponding reflectance value changes are used for the quantification of the two inorganic nitrogen species in the ranges 6.5-100.0 or 2.1-15.0 mg N L-1 for ammonium and 18.2-100.0 or 4.2-15.0 mg N L-1 for nitrate when BTB or NY are used, respectively. Under optimal conditions the limits of quantification of ammonium and nitrate in the case of BTB were determined as 6.5 and 18.2 mg N L-1, respectively, while the corresponding values in the case of NY were found to be 2.1 and 4.2 mg N L-1. The newly developed µPAD was stable for 62 days when stored in a freezer and 1 day at ambient temperature. It was validated with a certified reference material and successfully applied to the determination of ammonium and nitrate in spiked environmental water samples and soil extracts.

Temporal variability in nutrient concentrations and loads in the River Tamar and its catchment (SW England) between 1974 and 2004.

This study reports the results from the analyses of a 30-year (1974-2004) river water quality monitoring dataset for NO x -N (NO3-N + NO2-N), NH4-N, PO4-P and SiO2-Si at the tidal limit of the River Tamar (SW England), an agriculturally dominated and sparsely populated catchment. Annual mean concentrations of NH4-N, PO4-P and SiO2-Si were similar to other rural UK rivers, while annual mean concentrations of NO x -N were clearly lower. Estimated values for the 1940s were much lower than for those of post-1974, at least for NO3-N and PO4-P. Flow-weighted mean concentrations of PO4-P decreased by approximately 60 % between 1974 and 2004, although this change cannot be unequivocally ascribed to either PO4-P stripping from sewage treatment work effluents or reductions in phosphate fertiliser applications. Lower-resolution sampling (to once per month) in the late 1990s may also have led to the apparent decline; a similar trend was also seen for NH4-N. There were no temporal trends in the mean concentrations of NO x -N, emphasising the continuing difficulty in controlling diffuse pollution from agriculture. Concentrations of SiO2-Si and NO x -N were significantly and positively correlated with river flows ≤15 m(3) s(-1), showing that diffuse inputs from the catchment were important, particularly during the wet winter periods. In contrast, concentrations of PO4-P and NH4-N did not correlate across any flow window, despite the apparent importance of diffuse inputs for these constituents. This observation, coupled with the absence of a seasonal (monthly) cycle for these nutrients, indicates that, for PO4-P and NH4-N, there were no dominant sources and/or both undergo extensive within-catchment processing. Analyses of nutrient fluxes reveal net losses for NO3-N and SiO2-Si during the non-winter months; for NO3-N, this may be due to denitrification. Areal fluxes of NO x -N from the catchment were towards the higher end of the range for the UK, while NH4-N and PO4-P were closer to the lower end of the ranges for these nutrients. These data, taken together with information on sestonic chlorophyll a, suggest that water quality in the lower River Tamar is satisfactory with respect to nutrients. Analyses of these monitoring data, which were collected at considerable logistical and monetary cost, have revealed unique insights into the environmental behaviour of key nutrients within the Tamar catchment over a 30-year period.

Sedimentary pools of phosphorus in the eutrophic Tamar estuary (SW England).

The speciation of phosphorus in Tamar estuary sediment has been determined at eight stations from Gunnislake (riverine end-member) to Torpoint (marine end-member) using the SEDEX (sedimentary extraction) sequential extraction scheme. The seasonal variability of total phosphorus and each of the five fractions is discussed. Total phosphorus, in the range 26-51 micromol g(-1), was stored in the relatively labile fractions, particularly in the iron oxide fraction. The well oxygenated waters of the macrotidal Tamar estuary ensure that the surface sediment layer remains oxic, providing an effective barrier that retains dissolved reactive phosphorus (DRP) within the sedimentary iron oxide fraction and prevents phosphorus migration from the sediment to the water column. The exchangeable and organic phosphorus sedimentary pools, although small, are therefore the main potential sources of phosphorus release to the water column. In particular, this study shows that exchangeable phosphorus and dissolved organic phosphorus (DOP) in the pore water are intimately linked. Finally, the distribution of phosphorus in the estuary is strongly influenced by particle morphology, with most of the sedimentary phosphorus forms exhibiting significant positive correlations with the percent of each grain size.

Monitoring of ammonia in marine waters using a passive sampler with biofouling resistance and neural network-based calibration.

A biofouling resistant passive sampler for ammonia, where the semi-permeable barrier is a microporous hydrophobic gas-diffusion membrane, has been developed for the first time and successfully applied to determine the time-weighted average concentration of ammonia in estuarine and coastal waters for 7 days. Strategies to control biofouling of the membrane were investigated by covering it with either a copper mesh or a silver nanoparticle functionalised cotton mesh, with the former approach showing better performance. The effects of temperature, pH and salinity on the accumulation of ammonia in the newly developed passive sampler were studied and the first two parameters were found to influence it significantly. A universal calibration model for the passive sampler was developed using the Group Method Data Handling algorithm based on seawater samples spiked with known concentrations of total ammonia under conditions ranging from 10 to 30 °C, pH 7.8 to 8.2 and salinity 20 to 35. The newly developed passive sampler is affordable, user-friendly, reusable, sensitive, and can be used to detect concentrations lower than the recently proposed guideline value of 160 µg total NH3-N L-1, for a 99% species protection level, with the lowest concentration measured at 17 nM molecular NH3 (i.e., 8 µg total NH3-N L-1 at pH 8.0 and 20 °C). It was deployed at four field sites in the coastal waters of Nerm (Port Phillip Bay), Victoria, Australia. Good agreement was found between molecular ammonia concentrations obtained with passive and discrete grab sampling methods (relative difference, - 12% to - 19%).

Development of a micro-distillation microfluidic paper-based analytical device as a screening tool for total ammonia monitoring in freshwaters.

An easy-to-use, portable 3D microfluidic paper-based analytical device (µPAD) for the determination of total ammonia (i.e., ammonia + ammonium) in freshwaters is described. It consists of two layers of paper patterned with hydrophilic circular zones, one impregnated with sodium hydroxide (sample zone) and another (detection zone) with an acid-base indicator (nitrazine yellow (NY) or bromothymol blue (BTB)), separated by a µ-distillation chamber. Ammonium ions present in the water sample are converted into ammonia gas by reaction with sodium hydroxide in the sample zone. Ammonia then diffuses through a headspace and reacts with an acid-base indicator in the detection zone, the reflectance of which can be related to the total ammonia concentration. The analytical signal at 7.8 mg L-1 offered by the µ-distillation chamber-based µPAD is more than double of that obtained using a gas-permeable membrane. The proposed µPAD is characterised by a limit of detection of 0.32 or 0.47 mg N L-1 and working concentration ranges of 0.5-3.0 mg N L-1 or 2.0-10 mg N L-1 when using NY or BTB indicators, respectively. This is the first µPAD whose working range covers almost the entire trigger value range (0.32-2.3 mg N L-1) for ammonia nitrogen in freshwater systems which makes it suitable as a field screening tool for ammonia in freshwaters. An inter- and intra-device repeatability of 7.6% and 9.0%, respectively, has been achieved for the NY-based µPAD (3 mg N L-1) and 13 and 2.5% (8 mg N L-1) for the BTB-based µPAD. The NY-based µPAD is stable under vacuum for at least 12 days at room temperature, and 56 days if stored in a freezer (≤-20 °C).

Developments of microfluidic paper-based analytical devices (µPADs) for water analysis: A review.

Water pollution is a serious environmental problem affecting millions of people, and the demand for frequent water quality monitoring is increasing. The need for analytical platforms that combine high sensitivity, selectivity and accuracy with low cost, portability and user friendliness remains a challenge. Microfluidic paper-based analytical devices (µPADs) are recognised as a powerful analytical platform that can satisfy these requirements. The aim of this review is to provide a detailed overview of the µPADs that have been developed for the determination of important water quality parameters, such as nutrients, metals and organic contaminants, in a range of waters. A description of the fabrication and detection methods selected for these applications is provided, and the performance of the µPADs with respect to their precision and accuracy is critically assessed. The potential of these devices for real-life applications is also critically examined, particularly if they can determine the concentrations specified in water quality guidelines or the maximum recommended concentrations for various waters, as well as if they are suitable for field applications.

Gas-diffusion-based passive sampler for ammonia monitoring in marine waters.

A novel passive sampler based on gas-diffusion across a hydrophobic membrane is described for the determination of the time-weighted average concentration of dissolved molecular ammonia in high ionic strength aquatic environments, such as sea, coastal and estuarine waters, for a period of 3 days. The passive sampler developed is cheap, easy-to-use, reusable, and has a dynamic concentration range of 2.0-12µM, which covers the water quality guideline trigger value of 11.4µM (160µgL-1 NH3-N) for high conservation value waters, making this a powerful new tool for water quality managers involved in long-term ammonia monitoring. The gas-diffusion-based passive sampler was calibrated under laboratory conditions and deployed in a tank of seawater in the laboratory and at an estuarine site for proof of concept, and a good agreement between passive and spot sampling was achieved in both cases.

Colorimetric detection based on localised surface plasmon resonance of gold nanoparticles: Merits, inherent shortcomings and future prospects.

Localised surface plasmon resonance (LSPR) of gold nanoparticles (AuNPs) has been exploited for two decades in analytical science and has proven to be a powerful tool for the detection of various kinds of substances including small molecules, ions, macro biomolecules and microbes. Detection can be performed by visual colour change observations, photometry or resonance light scattering. A wide range of applications have been studied in the areas of environmental, pharmaceutical and biological analysis and clinical diagnosis. In this article, some fundamental aspects and important applications involving LSPR of AuNPs are reviewed. Several inherent shortcomings of these techniques and possible strategies to circumvent them are discussed.

The molybdenum blue reaction for the determination of orthophosphate revisited: Opening the black box.

The molybdenum blue reaction, used predominantly for the determination of orthophosphate in environmental waters, has been perpetually modified and re-optimised over the years, but this important reaction in analytical chemistry is usually treated as something of a 'black box' in the analytical literature. A large number of papers describe a wide variety of reaction conditions and apparently different products (as determined by UV-visible spectroscopy) but a discussion of the chemistry underlying this behaviour is often addressed superficially or not at all. This review aims to rationalise the findings of the many 'optimised' molybdenum blue methods in the literature, mainly for environmental waters, in terms of the underlying polyoxometallate chemistry and offers suggestions for the further enhancement of this time-honoured analytical reaction.

The use of on-line UV photoreduction in the flow analysis determination of dissolved reactive phosphate in natural waters.

A highly sensitive flow analysis manifold for rapid determination of dissolved reactive phosphate was developed which uses ethanol and UV light to reduce phosphomolybdic acid, instead of the reactive and short-lived chemical reductants typically employed in molybdenum blue chemistry. This reaction is impractical to perform reproducibly in batch mode, yet is very simple to handle in a flow analysis system and uses a single, very long-lived reagent solution. Interference from common inorganic anions and organic phosphorus species was minimal, and good spike recoveries for a range of sample matrices were obtained. The proposed flow analysis system is characterised by a limit of detection of 1.3 µg L(-1) P, linear range of 5-1000 µg L(-1) P, dynamic range of 5-5000 µg L(-1) P, repeatability of 0.8% (1000 µg L(-1) P, n=10) and 5.6% (10 µg L(-1) P, n=10), and sample throughput of 57 h(-1). It is expected that this method will improve the feasibility of autonomous long-term environmental monitoring of dissolved reactive phosphate using inexpensive apparatus.

The nature of the salt error in the Sn(II)-reduced molybdenum blue reaction for determination of dissolved reactive phosphorus in saline waters.

Sn(II) is a well-known reductant used in the formation of phosphomolybdenum blue for the determination of dissolved reactive phosphorus (DRP) in waters because it provides rapid and quantitative reduction. However, in saline waters, this method suffers from a salt error which causes a significant decrease in sensitivity. This phenomenon has never been adequately explained in the literature. The Murphy and Riley method, which uses Sb(III) and ascorbic acid for the reduction step, is preferred for DRP determination in saline waters because it is unaffected by salinity, but it exhibits a sensitivity approximately 30% lower than that when Sn(II) is used as the reductant without Cl(-) interference. This study investigates the processes causing the salt error and possible ways of minimizing it, so that the benefits of Sn(II) reduction on the molybdenum blue reaction rate and sensitivity may be exploited in the determination of low levels of DRP in marine and estuarine waters. It has been established that the salt error is caused by the formation of Sn(IV) chloro-complexes which compete with the formation of Sn(IV)-substituted phosphomolybdenum blue, forcing the reaction to proceed via the much slower, less favourable process of direct reduction that occurs in methods using organic reductants such as ascorbic acid.

Applications of everyday IT and communications devices in modern analytical chemistry: A review.

This paper reviews the development and recent use of everyday communications and IT equipment (mobile phones, digital cameras, scanners, webcams, etc) as detection devices for colorimetric chemistries. Such devices can readily be applied for visible detection using reaction formats such as microfluidic paper based analytical devices (µPADs), indicator papers, and well plate reaction vessels. Their use is highly advantageous with respect to cost, simplicity and portability, and offers many opportunities in the areas of point of care diagnosis, and at-site monitoring of environmental, agricultural, food and beverage parameters.

Seawater induced release and transformation of organic and inorganic phosphorus from river sediments.

This paper reports an investigation of the release of organic and inorganic phosphorus from a riverine sediment subjected to salinity conditions typical of estuarine mixing. Freshwater sediment was mixed with filtered river water in a thermostatted reactor, and allowed to equilibrate under aerobic conditions for 3 days. Salinity was then increased in a stepwise manner by addition of filtered low-nutrient seawater over a period of 4 days. A control experiment was performed in a second reactor by substituting ultrapure water for seawater. Using a flow injection analysis method for measurement of filterable reactive phosphorous (FRP, the so-called inorganic fraction) and filterable organic phosphorous (FOP), it was found that rapid releases of both FOP and FRP occurred at salinities of >/=10 per thousand. Over the 4-day experimental period, sediment release increased the filterable P concentration by approx. 50 microgL(-1), and of this, nearly half was initially present as FOP, which subsequently underwent rapid mineralisation to FRP. The observed behaviour may be explained by a combination of salinity induced plasmolysis of sediment bacteria and ion exchange by suspended sediment particles.

A novel technique for the pre-concentration and extraction of inositol hexakisphosphate from soil extracts with determination by phosphorus-31 nuclear magnetic resonance.

Inositol hexakisphosphate (IP6) is often the dominant form of soil organic phosphorus (P), but is rarely investigated because of the analytical difficulties encountered in its extraction, separation, and detection in environmental samples. In particular, recent advances in the study of soil organic P with 31P nuclear magnetic resonance (NMR) have been of limited use for the study of IP6, because the technique does not discriminate between IP6 and other forms of P. This was addressed by developing a novel analytical procedure using the retentive properties of gel-filtration gels for IP6, which allows the combined selective extraction and pre-concentration of IP6 from soil extracts with determination by 31P NMR. While the technique is still in the developmental stage, the results demonstrate that the gel does not interfere with 31P NMR analysis and retains IP6 to concentrations well above those required to give clear spectral signals. The technique has considerable potential for application to the study of IP6 in soil extracts and water samples and, with development, could help to answer fundamental questions regarding the dynamics of organic P in the environment.

The use of a polymer inclusion membrane for separation and preconcentration of orthophosphate in flow analysis.

A highly sensitive flow analysis system has been developed for the trace determination of reactive phosphate in natural waters, which uses a polymer inclusion membrane (PIM) with Aliquat 336 as the carrier for on-line analyte separation and preconcentration. The system operates under flow injection (FI) and continuous flow (CF) conditions. Under optimal FI conditions the system is characterised by a linear concentration range between 0.5 and 1000 µg L(-1)P, a sampling rate of 10h(-1), a limit of detection of 0.5 µgL(-1)P and RSDs of 3.2% (n = 10, 100 µg L(-1)) and 7.7% (n = 10, 10 µg L(-1)). Under CF conditions with 10 min stop-flow time and sample solution flow rate of 1.32 mL min(-1) the flow system offers a limit of detection of 0.04 µg L(-1)P, a sampling rate of 5h(-1) and an RSD of 3.4% (n=5, 2.0 µg L(-1)). Interference studies revealed that anions commonly found in natural waters did not interfere when in excess of at least one order of magnitude. The flow system, operating under CF conditions, was successfully applied to the analysis of natural water samples containing concentrations of phosphate in the low µg L(-1)P range, using the multipoint standard addition method.

Flow injection analysis as a tool for enhancing oceanographic nutrient measurements--a review.

Macronutrient elements (C, N and P) and micronutrient elements (Fe, Co, Cu, Zn and Mn) are widely measured in their various physico-chemical forms in open ocean, shelf sea, coastal and estuarine waters. These measurements help to elucidate the biogeochemical cycling of these elements in marine waters and highlight the ecological and socio-economic importance of the oceans. Due to the dynamic nature of marine waters in terms of chemical, biological and physical processes, it is advantageous to make these measurements in situ and in this regard flow injection analysis (FIA) provides a suitable shipboard platform. This review, therefore, discusses the role of FIA in the determination of macro- and micro-nutrient elements, with an emphasis on manifold design and detection strategies for the reliable shipboard determination of specific nutrient species. The application of various FIA manifolds to oceanographic nutrient determinations is discussed, with an emphasis on sensitivity, selectivity, high throughput analysis and suitability for underway analysis and depth profiles. Strategies for enhancing sensitivity and minimizing matrix effects, e.g. refractive index (schlieren) effects and the important role of uncertainty budgets in underpinning method validation and data quality are discussed in some detail.

A paper-based device for measurement of reactive phosphate in water.

The preparation and evaluation of a simple paper-based device for the determination of reactive phosphate in natural and soil waters based on the formation of phosphomolybdenum blue is reported. Hydrophilic reagent zones were defined by printing filter paper with a hydrophobic paper-sizing agent using an inkjet printer. The molybdate/potassium antimony(III)-tartrate reagent and ascorbic acid reductant merged together from adjacent hydrophilic zones after spotting the liquid sample onto the paper. The colour intensity of the phosphoantimonylmolybdenum blue complex produced was measured using a flat bed scanner. Under optimal conditions, the paper-based device is characterised by a working range of 0.2-10 mg L(-1) P, limits of detection and quantitation of 0.05 and 0.16 mg L(-1) P, respectively, and repeatability of less than 2% RSD at 1 and 5 mg L(-1) P (n=10 and 8). There was no significant difference between the results obtained using the paper-based device and reference methods (t=0.0135, P (T≤t) two-tail=0.9892, df=38). In its optimum form, the paper-based device was stable for up to 15 day under ambient conditions, and up to 122 day when stored at ≤-20 °C. The small dimensions, minimal reagent consumption, low cost (a few cents), ease of operation and favourable analytical performance make the proposed paper-based device attractive for on-site environmental monitoring and analysis.

Field measurement of nitrate in marine and estuarine waters with a flow analysis system utilizing on-line zinc reduction.

A sensitive reagent-injection flow analysis method for the spectrophotometric determination of nitrate in marine, estuarine and fresh water samples is described. The method is based on the reduction of nitrate in a micro column containing zinc granules at pH 6.5. The nitrite formed is reacted with sulfanilamide and N-(1-naphthyl)ethylene diamine (Griess reagent), and the resulting azo compound is quantified spectrophotometrically at 520 nm. Water samples in the range of 3-700 µg L(-1) NO(3)(-)-N can be processed with a throughput of up to 40 samples per hour, a detection limit of 1.3 µg L(-1) and reproducibility of 1.2% RSD (50 µg L(-1) NO(3)(-)-N, n=10). The proposed method was successfully applied for the determination of nitrate in estuarine waters and the reliability was assessed by the analyses of certified reference materials and recovery experiments. The method is suitable for waters with a wide range of salinities, and was successfully used for more than 3200 underway nitrate measurements aboard SV Pelican1 in the "Two Bays" cruise in January 2010.