Electrode, membrane: ammonia

While the majority of enzyme electrodes fabricated have been rather large devices, there have been some recent reports concerning the development of miniaturized and even microsensors. For example, MeyerhoflF (M5) prepared an essentially disposable urea sensor (tip diameter 3 mm) by immobilizing urease at the surface of a new type of polymer-membrane electrode-based ammonia sensor (see Fig. 4). Alexander and Joseph (Al) have also prepared a new miniature urea sensor by immobilizing urease at the surface of pH-sensitive antimony wire. Similarly, lannello and Ycynych (II) immobilized urease on a pH-sensitive iridium dioxide electrode. In these latter investigations, ammonia liberated from the enzyme-catalyzed reaction alters the pH in the thin film of enzyme adjacent to the pH-sensitive wire. 

M5. Meyerhoff, M. E., Preparation and response properties of selective bioelectrodes utilizing polymer membrane electrode-based ammonia gas sensors. Anal. Lett. 13,1345-1357 (1980). 

The resulting ammonium ions can be detected with a cation-sensitive glass membrane. Alternatively, one could use a gas-sensing electrode for ammonia in place of the glass electrode, so that interferences from H , Na", and K are reduced. 

A limiting factor on the speed of response of an enzyme-based electrode is the response time of the base electrode. Electrodes based on gas-permeable membranes (ammonia or carbon dioxide) have longer response times than those without such membranes (ISEs, pH glass electrodes, ISEETs). In both cases, the response time of the electrode is the limiting factor at low substrate concentration. Response times of the order of 10-30 s can easily be obtained for concentrations in the range 0.1-1 mmoll... 

The ammonia probe electrode has been designed for the measurement of ammonia concentrations in aqueous solutions and also the ammonium ion concentration once the sample has been converted to ammonia. The ammonia is detected by measuring the effect on the pH of an ammonium chloride solution, separated from the sample by a gas-permeable hydrophobic membrane (figure A.3). When the probe is in contact with the ammonia solution, the internal solution between the pH electrode membrane and the gas-permeable membrane gains or loses ammonia gas through the latter, until the partial pressure (activity) of ammonia is the same on both sides. Thus the pH of the internal solution is proportional to the free ammonia concentration in the sample. 

Ion-selective electrodes can also become sensors (qv) for gases such as carbon dioxide (qv), ammonia (qv), and hydrogen sulfide by isolating the gas in buffered solutions protected from the sample atmosphere by gas-permeable membranes. Typically, pH glass electrodes are used, but electrodes selective to carbonate or sulfide may be more selective. 

Many other cyclic and noncyclic organic carriers with remarkable ion selectivities have been used successfiilly as active hosts of various liquid membrane electrodes. These include the 14-crown-4-ether for lithium (30) 16-crown-5 derivatives for sodium bis-benzo-18-crown-6 ether for cesium the ionophore ETH 1001 [(R,R)-AA -bisd l-ethoxycarbonyl)undecyl-A,yVl-4,5-tctramcthyl-3,6-dioxaoctancdiamide] for calcium the natural macrocyclics nonactin and monensin for ammonia and sodium (31), respectively the ionophore ETH 1117 for magnesium calixarene derivatives for sodium (32) and macrocyclic thioethers for mercury and silver (33). 

A representative example of a biocatalytic membrane electrode is an electrode for L-arginine The bacterium streptococcus faecium is immobilized on the gas permeable membrane of an ammonia electrode. Arginine deiminase in the bacterium catalyzes the following reaction... 

The concept of a biocatalytic membrane electrode has been extended to the use of a tissue slice as the catalytic layer. An example of this approach is an electrode for AMP which consists of a slice of rabbit muscle adjacent to an ammonia gas electrode. NHj is produced by enzymatic action of rabbit muscle constituents on AMP The electrode exhibits a linear range of 1.4 x 10 to 1.0 x 10 M with a response time varying from 2.5 to 8.5 min, depending on the concentration. Electrode lifetime is about 28 days when stored between use in buffer with sodium azide to prevent bacterial growth. Excellent selectivity enables AMP to be determined in serum. 

Although rum ammonia levels are not routinely measured, it is a useful indicator of Reye s syndrome and should be monitored in newborns at risk of developing hyperammonemia Ammonia is produced in many analytically useful enzyme reactions and the ammonium ISE has been used as the base sensor in several enzyme electrodes (see next section). In addition to valinomycin, other antibiotics such as the nonactin homalogs and gramicidins also behave as ionophores. The nonactin homolo were originally studied for their ability to selectively bind potassiiun ions It was then discovered that ammonium ions were preferred over potassium ions, and the selectivity coefficient Knh+ = 0.12 was reported. Since ammonia is present at fairly low levels in serum, this selectivity is not sufficient to to accurately measure NH4 in the presence of K. An extra measure of selectivity can be gained by using a gas permeable membrane to separate the ammonia gas from the sample matrix... 

Therefore, the ISE potential depends on the CO2 partial pressure with Nernstian slope. Contemporary microporous hydrophobic membranes permitted the construction of a number of gas probes, developed mainly by the Orion Research Company (for a survey see [143]. The most important among these sensors is the ammonia electrode, in which ammonia diffusing through the membrane affects the pH at a glass electrode. Other electrodes based on similar principles respond to SO2, HCN, H2S (with an internal S ISE), etc. The ammonia probe has a better detection limit than the ammonium ion ISE based on the non-actin ionophore. The response time of gas probes depends mostly on the rate of diffusion of the test gas through the microporous medium [77,143]. 

Tissue electrodes [2, 3, 4, 5, 45,57], In these biosensors, a thin layer of tissue is attached to the internal sensor. The enzymic reactions taking place in the tissue liberate products sensed by the internal sensor. In the glutamine electrode [5, 45], a thick layer (about 0.05 mm) of porcine liver is used and in the adenosine-5 -monophosphate electrode [4], a layer of rabbit muscle tissue. In both cases, the ammonia gas probe is the indicator electrode. Various types of enzyme, bacterial and tissue electrodes were compared [2]. In an adenosine electrode a mixture of cells obtained from the outer (mucosal) side of a mouse small intestine was used [3j. The stability of all these electrodes increases in the presence of sodium azide in the solution that prevents bacterial decomposition of the tissue. In an electrode specific for the antidiuretic hormone [57], toad bladder is placed over the membrane of a sodium-sensitive glass electrode. In the presence of the antidiuretic hormone, sodium ions are transported through the bladder and the sodium electrode response depends on the hormone concentration. 

The bacteria consume oxygen during ammonia oxidation, so oxygen depletion can be detected by using an oxygen electrode. A combined creatinine sensor thus consists of a cellulose dialysis membrane, immobilized creatinine deaminase, immobilized nitrifying bacteria and an oxygen electrode [130]. 

Reflectance measurements provided an excellent means for building an ammonium ion sensor involving immobilization of a colorimetric acid-base indicator in the flow-cell depicted schematically in Fig. 3.38.C. The cell was furnished with a microporous PTFE membrane supported on the inner surface of the light window. The detection limit achieved was found to depend on the constant of the immobilized acid-base indicator used it was lO M for /7-Xylenol Blue (pAT, = 2.0). The response time was related to the ammonium ion concentration and ranged from 1 to 60 min. The sensor remained stable for over 6 months and was used to determine the analyte in real samples consisting of purified waste water, which was taken from a tank where the water was collected for release into the mimicipal waste water treatment plant. Since no significant interference fi-om acid compounds such as carbon dioxide or acetic acid was encountered, the sensor proved to be applicable to real samples after pH adjustment. The ammonium concentrations provided by the sensor were consistent with those obtained by ion chromatography, a spectrophotometric assay and an ammonia-selective electrode [269]. 

Figure 4.2 — (A) Schematic diagram of an ammonia-N-sensitive probe based on an Ir-MOS capacitor. (Reproduced from [20] with permission of the American Chemical Society). (B) Pneumato-amperometric flow-through cell (a) upper Plexiglas part (b) metallized Gore-Tec membrane (c) auxiliary Gore-Tec membrane (d) polyethylene spacer (e) bottom Plexiglas part (/) carrier stream inlet (g) carrier stream outlet. (C) Schematic representation of the pneumato-amperometric process. The volatile species Y in the carrier stream diffuses through the membrane pores to the porous electrode surface in the electrochemical cell and is oxidized or reduced. (Reproduced from [21] with permission of the American Chemical Society).

Figure 4.15 — (A) Tubular flow-through electrode 1 Perspex body 2 conducting epoxy cylinder 3 mobile carrier PVC membrane 4 electric cable 5 channel (1.2 mm ID) 6 holders 7 screws 8 0-rings. (B) Schematic diagram of a system for on-line monitoring of ammonia ISE tubular flow-through ammonium ion-selective electrode R reference electrode W waste. (Reproduced from [137] with permission of the Royal Society of Chemistry).

Figure 5.7 shows a typical application of gas-diffusion membranes isolation of the circulating sample from a voltammetric or potentiometric electrode for the electrochemical determination of gaseous species. The ion-selective electrode depicted in this Figure includes a polymer membrane containing nonactin that is used for the potentiometric determination of ammonia produced in biocatalytic reactions. Interferences from alkali metal ions are overcome by covering the nonactin membrane with an outer hydro-... 

Several combined electrodes have been developed to measure dissolved gases such as carbon dioxide (CO2), ammonia (NH3) and sulphur dioxide (S02). These sensor-like electrodes contain an internal solution that is isolated from the external solution by an impermeable membrane. The membrane forbids the passage of water or ions but is transparent to gas molecules dissolved in solution (Fig. 18.4). 

Immobilized enzyme (wide range) membranes with oxygen, ammonia, or carbon dioxide electrodes Universal Sensors, USA... 

Use an ammonia electrode (Orion Model 95-10, Beckman Model 39565 or equivalent) along with a readout device, such as a pH meter with expanded millivolt scale between -700 mV and +700 mV or a specific ion meter. The electrode assembly consists of a sensor glass electrode and a reference electrode mounted behind a hydrophobic gas-permeable membrane. The membrane separates the aqueous sample from an ammonium chloride internal solution. Before analysis, the sample is treated with caustic soda to convert any NH4+ ion present in the sample into NH3. The dissolved NH3 in the sample diffuses through the membrane until the partial pressure of NH3 in the sample becomes equal to that in the internal solution. The partial pressure of ammonia is proportional to its concentration in the sample. The diffusion of NH3 into the internal solution increases its pH, which is measured by a pH electrode. The chloride level in the internal standard solution remains constant. It is sensed by a chloride ion-selective electrode which serves as the reference electrode. 

Another example may be checking of the blank response of an ammonia ion selective electrode under different experimental conditions, namely the age of the sensor membrane and the speed at which the measuring solution is stirred. 

The samples for on-line analysis were taken through a polysulfone membrane [37]. The seven channel automatic analyzer system consisted of six air-segmented continuous flow-wet chemical analyzers (Skalar analytics) to measure the concentration of ammonia with ion selective electrode (Philips IS-570), phosphate with ammonium molybdate at 880 nm, reducing sugar with pHBAH at 410 nm, methionine with Na-nitroprusside at 505 nm, cephalosporin C with... 

Enzyme-selective electrodes (Fig. 17.11) have been made as a membrane containing immobilized enzymes placed over a pH electrode or over a gas electrode such as an ammonia electrode for potentiometric detection, or over an oxygen electrode for amperometric detection. The products of the reaction of enzyme with substrate are detected by the electrode. 

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