Layered Architectural Fabrication of a Novel Sulfite Nanobiosensor by Encapsulation of Sulfite Oxidase on a Polypyrrole-Multiwalled Carbon Nanotubes Composite Decorated with Platinum Nanoparticles.

The fabrication of a highly selective and ultrasensitive sulfite nanobiosensor based on a layered architectural fabrication aided by the encapsulation of sulfite oxidase (SOx) in Nafion (Naf) matrix on a multiwalled carbon nanotubes-polypyrrole (MWCNTs-PPy) composite decorated with platinum nanoparticles (PtNPs) is described. The MWCNTs are deposited in the inner layer on a Pt electrode during electropolymerization of pyrrole (Py), followed by decoration with a PtNPs layer and subsequent encapsulation of SOx with Naf in the third layer capped with a fourth thin PtNPs layer. Images obtained by field emission scanning electron microscopy (FESEM) reveal that high-density PtNPs are deposited onto the 3D nanostructured inner MWCNTs-PPy layer and the electrochemical behavior is investigated. A large surface area provided by the incorporation of MWCNTs in the composite and decoration with PtNPs enables increased SOx loading, SOx retention, and substantial improvement in sensing performance. The resulting layered PtNPs/SOx-Naf/PtNPs/MWCNTs-PPy nanobiosensor exhibits a fast response time (within 3 s), a linear calibration range of 20 nmm - 6 m with an excellent sensitivity of 71 µA mm-1  cm-2 and a detection limit of 5.4 nm. The nanobiosensor  was effective in discriminating against common interferants and  was successfully applied to sulfite determination in real samples.

Rolling chain amplification based signal-enhanced electrochemical aptasensor for ultrasensitive detection of ochratoxin A.

A novel electrochemical aptasensor is described for rapid and ultrasensitive detection of ochratoxin A (OTA) based on signal enhancement with rolling circle amplification (RCA). The primer for RCA was designed to compose of a two-part sequence, one part of the aptamer sequence directed against OTA while the other part was complementary to the capture probe on the electrode surface. In the presence of target OTA, the primer, originally hybridized with the RCA padlock, is replaced to combine with OTA. This induces the inhibition of RCA and decreases the OTA sensing signal obtained with the electrochemical aptasensor. Under the optimized conditions, ultrasensitive detection of OTA was achieved with a limit of detection (LOD) of 0.065 ppt (pg/mL), which is much lower than previously reported. The electrochemical aptasensor was also successfully applied to the determination of OTA in wine samples. This ultrasensitive electrochemical aptasensor is of great practical importance in food safety and could be widely extended to the detection of other toxins by replacing the sequence of the recognition aptamer.

Biofunctionalisation of Polypyrrole Nanowires Array with Sulfite Oxidase Coupled with the Integration of Platinum Nanoparticles for Ultrasensitive Amperometric Detection of Sulfite.

Sulfite determination in foods and alcoholic beverages is a common requirement by food and drug administration organisations in most countries. In this study, the enzyme, sulfite oxidase (SOx), is used to biofunctionalise a platinum-nanoparticle-modified polypyrrole nanowire array (PPyNWA) for the ultrasensitive amperometric detection of sulfite. A dual-step anodisation method was used to prepare the anodic aluminum oxide membrane used as a template for the initial fabrication of the PPyNWA. PtNPs were subsequently deposited on the PPyNWA by potential cycling in a platinum solution. The resulting PPyNWA-PtNP electrode was then biofuntionalised by adsorption of SOx onto the surface. The confirmation of the adsorption of SOx and the presence of PtNPs in the PPyNWA-PtNPs-SOx biosensor was verified by scanning electron microscopy and electron dispersive X-ray spectroscopy. Cyclic voltammetry and amperometric measurements were used to investigate the properties of the nanobiosensor and to optimise its use for sulfite detection. Ultrasensitive detection of sulfite with the PPyNWA-PtNPs-SOx nanobiosensor was accomplished by use of 0.3 M pyrrole, 10 U mL-1 of SOx, adsorption time of 8 h, a polymerisation period of 900 s, and an applied current density of 0.7 mA cm-2. The response time of the nanobiosensor was 2 s, and its excellent analytical performance was substantiated with a sensitivity of 57.33 µA cm-2 mM-1, a limit of detection of 12.35 nM, and a linear response range from 0.12 to 1200 µM. Application of the nanobiosensor to sulfite determination in beer and wine samples was achieved with a recovery efficiency of 97-103%.

Towards commercial scale postcombustion capture of CO2 with monoethanolamine solvent: key considerations for solvent management and environmental impacts.

Chemical absorption with aqueous amine solvents is the most advanced technology for postcombustion capture (PCC) of CO(2) from coal-fired power stations and a number of pilot scale programs are evaluating novel solvents, optimizing energy efficiency, and validating engineering models. This review demonstrates that the development of commercial scale PCC also requires effective solvent management guidelines to ensure minimization of potential technical and environmental risks. Furthermore, the review reveals that while solvent degradation has been identified as a key source of solvent consumption in laboratory scale studies, it has not been validated at pilot scale. Yet this is crucial as solvent degradation products, such as organic acids, can increase corrosivity and reduce the CO(2) absorption capacity of the solvent. It also highlights the need for the development of corrosion and solvent reclamation technologies, as well as strategies to minimize emissions of solvent and degradation products, such as ammonia, aldehydes, nitrosamines and nitramines, to the atmosphere from commercial scale PCC. Inevitably, responsible management of aqueous and solid waste will require more serious consideration. This will ultimately require effective waste management practices validated at pilot scale to minimize the likelihood of adverse human and environmental impacts from commercial scale PCC.

Galvanostatic entrapment of penicillinase into polytyramine films and its utilization for the potentiometric determination of penicillin.

A sensitive and reliable potentiometric biosensor for determination of penicillin has been developed by exploiting the self-limiting growth of the non-conducting polymer, polytyramine. Optimum polytyramine-penicillinase (PTy-PNCnase) films for potentiometric detection of penicillin were accomplished with monomer solutions which contained 0.03 M tyramine, 37 U/mL penicillinase, 0.01 M KNO3, and 3 mM penicillin with an applied current density of 0.8 mA/cm2 and an electropolymerisation time of 40 seconds. The potentiometric biosensor gave a linear concentration range of 3-283 µM for penicillin and achieved a minimum detectable concentration of 0.3 µM. The biosensor was successfully utilized for the detection of Amoxycillin and gave an average percentage recovery of 102±6%. Satisfactory recoveries of penicillin G were also achieved in milk samples with the potentiometric biosensor when concentrations are ≥20 ppm.

Vapor generation atomic absorption spectrometric determination of cadmium in environmental samples with in-line anion exchange separation.

A novel in-line anion exchange separation method is described for the removal of Mn(2+), Fe(3+), Ni(2+), Co(2+), Cu(2+), Zn(2+), and Pb(2+) from Cd(2+) and for its subsequent determination by vapor generation atomic absorption spectrometry. High Cd(2+) retention efficiency and maximum exclusion of other metal ions were achieved by using Cl(-), Br(-), or I(-) loaded strong base anion exchange resin and fiber in chloride medium. SO(4)(2-) and NO(3)(-) affected the retention of Cd(2+) on Cl(-) or Br(-) loaded exchangers but were beneficial for elution from I(-) loaded exchangers. Fast and efficient elution from the exchangers was achieved by using ethylenediamine solution. The removal of Zn(2+) was unnecessary as it prevents a decline in the sensitivity of cadmium response when ethylenediamine and high acid concentration were used. Under optimum conditions, the achievable detection limits (3sigma) with sample loadings of 0.48 and 9.6 mL were 40 ng L(-1) and 3 ng L(-1), respectively. Interference of residual organic matrix, in sample digests, with the separation and retention of cadmium were eliminated by use of microwave assisted digestion. The method was successfully applied to the determination of Cd in river sediment, fish liver, and water samples.

Fabrication of a novel DNA affinity biosensor based on hybridisation induced current by electrostatic repulsion of silicotungstic acid as a redox indicator.

We report on a novel DNA affinity biosensor which utilises the capture of a neutral charged single stranded (ss) morpholino DNA on a gold electrode to trigger an electrostatic repulsion of negatively charged silicotungstate anions and, in turn, enabled detection of the hybridisation of complementary base pairs. The repulsion of the anions, as a redox indicator, is reflected by a decrease in its electrochemical response with increasing target ss-DNA concentration. A theoretical framework for DNA detection by the affinity biosensor is proposed and verified by electrochemical measurements in the presence of the target ss-DNA by either dc cyclic voltammetry or Fourier transformed alternating current voltammetry (FTACV). The optimised conditions for the capture of the target ss-DNA and the electrochemical detection include 1 µM thiolated neutral morpholino oligo-nucleotide probe, hybridisation time of 10 min, 0.25 mM [α-SiW12O40]4-, and 25 mM phosphate buffer. In addition, the use of the 5th harmonic component of the FTACV gave the most sensitive response for the detection of the target ss-DNA. Under these conditions, the DNA affinity biosensor, based on FTACV detection, achieved a minimum detectable concentration of 0.1 pM ss-DNA and a linear concentration range of 0.1-1000 pM. The biosensor also successfully distinguished between some matched and mismatched base pairs.

A single step electrochemical integration of gold nanoparticles, cholesterol oxidase, cholesterol esterase and mediator with polypyrrole films for fabrication of free and total cholesterol nanobiosensors.

The development of free and total cholesterol nanobiosensors based on a single step electrochemical integration of gold nanoparticles (AuNPs), cholesterol oxidase (COx), cholesterol esterase (CE) and a mediator with polypyrrole (PPy) films is described. The incorporation of the various components in the PPy films was confirmed by chronopotentiometry, cyclic voltammetry (CV), scanning electron microscopy, energy dispersive X-ray analysis (SEM-EDX), and Fourier transformed infrared (FTIR) spectroscopy. The free cholesterol, PPy-NO3--Fe(CN)64--AuNPs-COx, nanobiosensor achieved a minimum detectable concentration of 5 µM, a linear concentration range of 5-25 µM and a sensitivity of 1.6 µA cm-2 µM-1 in 0.05 M phosphate buffer (pH 7.00). For the total cholesterol, PPy-NO3--Fe(CN)64--AuNPs-COx-CE, nanobiosensor which also involved the co-incorporation of cholesterol esterase (CE) with the other components, the achieved performances include a minimum detectable total cholesterol concentration of 25 µM, a broader linear concentration range of 25-170 µM and a lower sensitivity of 0.1 µA µM-1 cm-2. Owing to its high selectivity, the presence of common interferants did not affect the total cholesterol measurement with the PPy-NO3--Fe(CN)64--AuNPs-COx-CE nanobiosensor. Both nanobiosensors were successfully used for direct and indirect determination of total cholesterol in human blood serum samples.

Amperometric detection of glucose in fruit juices with polypyrrole-based biosensor with an integrated permselective layer for exclusion of interferences.

A novel polypyrrole (PPy)-based bilayer amperometric glucose biosensor integrated with a permselective layer has been developed for detection of glucose in the presence of interferences. It comprises of a PPy-GOx film grown, in the absence of electrolyte, as an inner layer, and a permselective PPy-Cl film as an outer layer. The PPy-GOx/PPy-Cl bilayer biosensor was effective in rejecting 98% of ascorbic acid and 100% of glycine, glutamic acid and uric acid. With an outer layer thickness of 6.6nm, the bilayer biosensor gave nearly identical glucose response to that of a single layer PPy-GOx biosensor. The biosensor also exhibited good reproducibility (1.9% rsd, n=10), high stability (more than 2months), wide linear range (0.5-24mM), low Km (8.4mM), high Imax (77.2µAcm-2), low detection limit (26.9µM) and good sensitivity (3.5µAcm-2mM-1). The bilayer biosensor was successfully employed for glucose determination in various fruit juices.

Recent advances in the development and utilization of modern anode materials for high performance microbial fuel cells.

Microbial fuel cells (MFCs) are novel bio-electrochemical device for spontaneous or single step conversion of biomass into electricity, based on the use of metabolic activity of bacteria. The design and use of MFCs has attracted considerable interests because of the potential new opportunities they offer for sustainable production of energy from biodegradable and reused waste materials. However, the associated slow microbial kinetics and costly construction materials has limited a much wider commercial use of the technology. In the past ten years, there has been significant new developments in MFCs which has resulted in several-fold increase in achievable power density. Yet, there is still considerable possibility for further improvement in performance and development of new cost effective materials. This paper comprehensively reviews recent advances in the construction and utilization of novel anodes for MFCs. In particular, it highlights some of the critical roles and functions of anodes in MFCs, strategies available for improving surface areas of anodes, dominant performance of stainless-steel based anode materials, and the emerging benefits of inclusion of nanomaterials. The review also demonstrates that some of the materials are very promising for large scale MFC applications and are likely to replace conventional anodes for the development of next generation MFC systems. The hurdles to the development of commercial MFC technology are also discussed. Furthermore, the future directions in the design and selection of materials for construction and utilization of MFC anodes are highlighted.

Rapid amperometric detection of trace metals by inhibition of an ultrathin polypyrrole-based glucose biosensor.

A sensitive and reliable inhibitive amperometric glucose biosensor is described for rapid trace metal determination. The biosensor utilises a conductive ultrathin (55 nm thick) polypyrrole (PPy) film for entrapment of glucose oxidase (GOx) to permit rapid inhibition of GOx activity in the ultrathin film upon exposure to trace metals, resulting in reduced glucose amperometric response. The biosensor demonstrates a relatively fast response time of 20s and does not require incubation. Furthermore, a complete recovery of GOx activity in the ultrathin PPy-GOx biosensor is quickly achieved by washing in 2mM EDTA for only 10s. The minimum detectable concentrations achieved with the biosensor for Hg(2+), Cu(2+), Pb(2+) and Cd(2+) by inhibitive amperometric detection are 0.48, 1.5, 1.6 and 4.0 µM, respectively. Also, suitable linear concentration ranges were achieved from 0.48-3.3 µM for Hg(2+), 1.5-10 µM for Cu(2+), 1.6-7.7 µM for Pb(2+) and 4-26 µM for Cd(2+). The use of Dixon and Cornish-Bowden plots revealed that the suppressive effects observed with Hg(2+) and Cu(2+) were via non-competitive inhibition, while those of Pb(2+) and Cd(2+) were due to mixed and competitive inhibition. The stronger inhibition exhibited by the trace metals on GOx activity in the ultrathin PPy-GOx film was also confirmed by the low inhibition constant obtained from this analysis. The biosensor was successfully applied to the determination of trace metals in tap water samples.

Nitrate biosensors and biological methods for nitrate determination.

The inorganic nitrate (NO3‾) anion is present under a variety of both natural and artificial environmental conditions. Nitrate is ubiquitous within the environment, food, industrial and physiological systems and is mostly present as hydrated anion of a corresponding dissolved salt. Due to the significant environmental and toxicological effects of nitrate, its determination and monitoring in environmental and industrial waters are often necessary. A wide range of analytical techniques are available for nitrate determination in various sample matrices. This review discusses biosensors available for nitrate determination using the enzyme nitrate reductase (NaR). We conclude that nitrate determination using biosensors is an excellent non-toxic alternative to all other available analytical methods. Over the last fifteen years biosensing technology for nitrate analysis has progressed very well, however, there is a need to expedite the development of nitrate biosensors as a suitable alternative to non-enzymatic techniques through the use of different polymers, nanostructures, mediators and strategies to overcome oxygen interference.

Inhibitive potentiometric detection of trace metals with ultrathin polypyrrole glucose oxidase biosensor.

A method, based on the inhibition of an ultrathin polypyrrole-glucose oxidase (PPy-GOx) potentiometric biosensor response, is described for the detection of Cu(2+), Hg(2+), Cd(2+) and Pb(2+) ions. Based on experimental conditions (0.2 M pyrrole, 500 U mL(-1) GOx, and an applied current density of 0.05 mA cm(-2) and a polymerization period of 500s) previously published by us, PPy-GOx films of approximately 55 nm thick were used to demonstrate the inhibitive potentiometric detection of selected trace metals down to 0.079 µM Cu(2+), 0.025 µM Hg(2+), 0.024 µM Pb(2+) and 0.044 µM Cd(2+). Furthermore, good linear concentration ranges were achieved for Cu(2+) (0.079-16 µM), Hg(2+) (0.025-5 µM), Pb(2+) (0.10-15 µM) and Cd(2+) (0.04-62 µM). The analysis of the nature of the inhibition of glucose oxidase in the PPy-GOx biosensor by these metals was achieved by Dixon and Cornish-Bowden plots. The shapes of the curves (exponential decay, parabolic and linear) obtained for the inhibitors suggest that the inhibition by the metal ions may not be exclusively directed at the essential -SH group, but involve additional binding sites of the enzyme. Dixon and Cornish-Bowden plots suggest that the inhibition is competitive for Cd(2+), while non-competitive inhibition was observed for other metal ions. The ultra-thin PPy-GOx film enabled improved permeability to the metal inhibitors than possible with conventional biosensors with thicker films and, hence, better reflects the actual inhibition effect of the trace metals on the enzyme activity. The use of the ultra-thin film also eliminated the usual need for incubation of the enzyme electrode for a long period in the presence of the inhibitors. Furthermore, a rapid recovery of the enzyme activity was achieved by simply washing the electrode with water and storing in phosphate buffer for 10-15 min. The proposed biosensing approach was successfully used for the detection of individual trace metals in tap water, achieving a 98-101% recovery.

Coupling of non-selective adsorption with selective elution for novel in-line separation and detection of cadmium by vapour generation atomic absorption spectrometry.

Non-selective adsorption of Cd(2+) ions on a cation exchange fiber and subsequent selective elution with a KI solution has been strategically utilized to develop a highly selective in-line separation of Cd(2+) ions from other metal ions for its rapid and reliable quantification by cold vapour-atomic absorption spectrometry. After retention of Cd(2+) with a high efficiency on cation exchange fiber, selective elution of the retained Cd(2+) was subsequently accomplished with 0.3M KI. Vapour generation of Cd for in-line CV-AAS determination was then achieved by merging the eluate with HCl and NaBH4. Interferences from most base metals with the vapour generation of Cd were eliminated by this approach, with the exception of Pb(2+)ions which was removed by co-precipitation with BaSO4 prior to the in-line separation. Substantial improvement in sensitivity of the in-line CV-AAS determination of Cd was achieved by increasing the sample loading time. A detection limit of 0.6 ng L(-1) (3σ) was obtained with sample loading time of 120 s, corresponding to a consumption of 24 mL of sample solution. Application of the method to the determination of Cd in certified sediment and fish samples gave a good agreement with the certified values. Further validation by recovery study in real fish sample digests and water gave average Cd recoveries of 98.7±1.0% for fish and 92±3% for water with RSD of 1.5% for fish and 4% for water, respectively.

A novel ultrasensitive phosphate amperometric nanobiosensor based on the integration of pyruvate oxidase with highly ordered gold nanowires array.

A novel phosphate amperometric nanobiosensor, based on an intimate integration of pyruvate oxidase (PyOx) and its cofactors, thiamine pyrophosphate (TPP) and flavin adenine dinucleotide (FAD), with a highly ordered gold nanowires array (AuNWA) has been developed. The successful integration of PyOx and the co-factors, via crosslinking with bovine serum albumin (BSA) and glutaraldehyde (GLA), onto the AuNWA was confirmed by cyclic voltammetry and amperometry. The resulting nanobiosensor achieved a detection limit of 0.1 µM, a linear concentration range of 12.5-1000 µM, and a sensitivity of 140.3 µA mM(-1)cm(-2). Notably, the incorporation of the AuNWA reduced the required PyOx concentration by 70-120 fold and the presence of common interferants, such as chloride, sulfate, fluoride, nitrite and nitrate ions did not interfere with phosphate detection. Furthermore, the nanobiosensor demonstrated a very high stability with repeated use over two weeks and was successfully used for the determination of phosphate in water samples with an average recovery of 96.6 ± 4.9%.

A novel GMO biosensor for rapid ultrasensitive and simultaneous detection of multiple DNA components in GMO products.

Since the introduction of genetically modified organisms (GMOs), there has been on-going and continuous concern and debates on the commercialization of products derived from GMOs. There is an urgent need for development of highly efficient analytical methods for rapid and high throughput screening of GMOs components, as required for appropriate labeling of GMO-derived foods, as well as for on-site inspection and import/export quarantine. In this study, we describe, for the first time, a multi-labeling based electrochemical biosensor for simultaneous detection of multiple DNA components of GMO products on the same sensing interface. Two-round signal amplification was applied by using both an exonuclease enzyme catalytic reaction and gold nanoparticle-based bio-barcode related strategies, respectively. Simultaneous multiple detections of different DNA components of GMOs were successfully achieved with satisfied sensitivity using this electrochemical biosensor. Furthermore, the robustness and effectiveness of the proposed approach was successfully demonstrated by application to various GMO products, including locally obtained and confirmed commercial GMO seeds and transgenetic plants. The proposed electrochemical biosensor demonstrated unique merits that promise to gain more interest in its use for rapid and on-site simultaneous multiple screening of different components of GMO products.

Integrated platform with magnetic purification and rolling circular amplification for sensitive fluorescent detection of ochratoxin A.

In this article, we report the detection of ochratoxin A (OTA) with excellent sensitivity with the two-aspect signal amplification treatments. Combining the unique property of magnetic nanoparticles and the high efficiency of the in vitro amplification of rolling circular amplification (RCA), the competitive sensing protocol for ultrasensitive detection of OTA was achieved in about 80 min. The excellent magnetic separation treatment could effectively avoid the interference of background fluorescent noise in the sensing system while the RCA could tremendously increase the hybridization sequence for the quantum dot labeled probes and further increase the sensing response signal. Afterwards, two factors affecting the final detection limit, concentration of RCA components and RCA reaction time, were all systematically optimized for the best sensing performance. The response of the optimized protocol for OTA detection is highly linear over the wider range from 10(-3) to 10 ppb, which is 3 orders improvement in sensing range, and the limit of detection is calculated to be as low as 0.13 ppt, which is 10,000 folds improvement compared with the traditional methods. More importantly, given the selected aptamer, this universal signal amplification protocol could be widely applied to other fields by just change the recognition sequence of the aptamer.

Integration of a highly ordered gold nanowires array with glucose oxidase for ultra-sensitive glucose detection.

A highly sensitive amperometric nanobiosensor has been developed by integration of glucose oxidase (GO(x)) with a gold nanowires array (AuNWA) by cross-linking with a mixture of glutaraldehyde (GLA) and bovine serum albumin (BSA). An initial investigation of the morphology of the synthesized AuNWA by field emission scanning electron microscopy (FESEM) and field emission transmission electron microscopy (FETEM) revealed that the nanowires array was highly ordered with rough surface, and the electrochemical features of the AuNWA with/without modification were also investigated. The integrated AuNWA-BSA-GLA-GO(x) nanobiosensor with Nafion membrane gave a very high sensitivity of 298.2 µA cm(-2) mM(-1) for amperometric detection of glucose, while also achieving a low detection limit of 0.1 µM, and a wide linear range of 5-6000 µM. Furthermore, the nanobiosensor exhibited excellent anti-interference ability towards uric acid (UA) and ascorbic acid (AA) with the aid of Nafion membrane, and the results obtained for the analysis of human blood serum indicated that the device is capable of glucose detection in real samples.

BiOI/TiO2 nanotube arrays, a unique flake-tube structured p-n junction with remarkable visible-light photoelectrocatalytic performance and stability.

A series of unique flake-tube structured p-n heterojunctions of BiOI/TiO2 nanotube arrays (TNTAs) were successfully prepared by loading large amounts of BiOI nanoflakes onto both the outer and inner walls of well-separated TiO2 nanotubes using anodization followed by the sequential chemical bath deposition (S-CBD) method. The as-prepared BiOI/TNTAs samples were characterized by X-ray diffraction, electron microscopy, X-ray photoelectron spectroscopy, UV-vis diffuse reflectance spectroscopy and nitrogen sorption. The photoelectrocatalytic (PEC) activity and stability of the BiOI/TNTAs samples toward degradation of methyl orange (MO) solutions under visible-light irradiation (λ > 420 nm) were evaluated. The visible-light PEC performance of BiOI/TNTAs samples was further confirmed by the transient photocurrent response test. The results from the current study revealed that the 5-BiOI/TNTAs sample exhibited the best PEC activity, favourable stability, and the highest photocurrent density among all the BiOI/TNTAs heterostructured samples. The combined effects of several factors may contribute to the remarkable visible-light PEC performance for the 5-BiOI/TNTAs sample including a 3D connected intertube spacing system and an open tube-mouth structure, strong visible-light absorption by BiOI, the formation of a p-n junction, larger specific surface area, and the impact of the applied external electrostatic field.

Progress and recent advances in phosphate sensors: a review.

This review covers the progress made in the development of sensors for inorganic and organic phosphates that are significant pollutants within the environmental and biological systems. Phosphate sensors in the forms of amperometric, potentiometric enzyme electrodes, plant tissue electrode and screen printed electrode are described. Instrumental probes such as fluorescence, chemiluminescence, luminescence and potentiometric ion selective electrodes are also described. Recent efforts on the use of voltammetric, potentiometric and amperometric biosensors for the determination of phosphate are highlighted.