Biosensor response time

The response time is often governed by the immobilized biocatalyst layer rather than by the sensor response, and there are many factors that affect the biosensor response time, as shown in Table 17.5. 

A bioelectrode functioning optimally has a short response time, which is often controlled by the thickness of the immmobilized enzyme layer rather than by the sensor as well as many other factors (see Table 7). The biosensor response time depends on (1) how rapidly the substrate diffuses through the solution to the membrane surface, (2) how rapidly the substrate diffuses through the membrane cmd reacts with the biocatalyst at the active site, and (3) how rapidly the products formed diffuse to the electrode surface where they are measured. Mathematical models describing this effea are thoroughly presented in the biosensor literature (5, 68). 

Generally, the thinner the membranes, the shorter is the biosensor response time. 

Simulation of the biosensor action usually involves calculation of the concentrations of the compounds as well as the response for the time interval from the beginning of the action up to the moment called the biosensor response time. The moment of the measurement depends on the type of the device. Devices operating in the stationary mode usually use the time when the absolute response, e.g., anodic or cathodic current, slope value falls below a given small value. Since the biosensor responses usually vary even in orders of magnitude, the response is normalized. In other words, the time Tneeded to achieve a given dimensionless decay rate e is accepted as the response time ... 

Use of aerosols — A large number of applications require biosensors with a rapid response. Exaiiq)les include alarm systems, in which urgency is vital, and flow injection analysis (FIA), in which a rapid response can increase the sample throughput and reduce the cost of analysis. Biosensor response times are very dependent on the thickness of the active layer (see 4.2.1). Aerosol vaporization of dissolved compounds deposits films that are thin and homogeneous. 

The low detection limit, high sensitivity, and fast response times of chemoreceptor-based biosensors result primarily from the extremely high binding constants of the receptor R for the target substrate S. The receptor—substrate binding may be described... 

The dye is excited by light suppHed through the optical fiber (see Fiber optics), and its fluorescence monitored, also via the optical fiber. Because molecular oxygen, O2, quenches the fluorescence of the dyes employed, the iatensity of the fluorescence is related to the concentration of O2 at the surface of the optical fiber. Any glucose present ia the test solution reduces the local O2 concentration because of the immobilized enzyme resulting ia an iacrease ia fluorescence iatensity. This biosensor has a detection limit for glucose of approximately 100 ]lM , response times are on the order of a miaute. 

While planar optical sensors exist in various forms, the focus of this chapter has been on planar waveguide-based platforms that employ evanescent wave effects as the basis for sensing. The advantages of evanescent wave interrogation of thin film optical sensors have been discussed for both optical absorption and fluorescence-based sensors. These include the ability to increase device sensitivity without adversely affecting response time in the case of absorption-based platforms and the surface-specific excitation of fluorescence for optical biosensors, the latter being made possible by the tuneable nature of the evanescent field penetration depth. 

CNTs offer an exciting possibility for developing ultrasensitive electrochemical biosensors because of their unique electrical properties and biocompatible nanostructures. Luong et al. have fabricated a glucose biosensor based on the immobilization of GOx on CNTs solubilized in 3-aminopropyltriethoxysilane (APTES). The as-prepared CNT-based biosensor using a carbon fiber has achieved a picoamperometric response current with the response time of less than 5 s and a detection limit of 5-10 pM [109], When Nation is used to solubilize CNTs and combine with platinum nanoparticles, it displays strong interactions with Pt nanoparticles to form a network that connects Pt nanoparticles to the electrode surface. The Pt-CNT nanohybrid-based glucose biosensor... 

Among organic constituent measurements, that of aggregate properties (BOD and COD) and specific parameters (TOC for example) has been well developed for more than 20 years. Concerning BOD, a recent review on biosensors [33] has been published. BOD biofilm-based sensors as well as respirometric systems, other measuring principles, and the commercial BOD instruments are discussed and compared regarding their performance characteristics like linearity, response time, precision, agreement between BOD values obtained from the biosensors and the conventional 5-day test, as well as toxic resistance to various compounds and operational stability. 

The method of enzyme immobilization constitutes a key factor in the construction of these systems as it is the biocatalytic membrane that largely determines sensitivity, stability and response-time characteristics of the biosensor. 

Table 3.2 lists the most salient microbial sensors reported to date, together with the type of immobilized microorganism and measurement used, and the response time and dynamic range achieved in each instance. As can be seen, most of these biosensors rely on amperometric measurements. Some of them are described in detail below. 

Biochemical oxygen demand (BOD) is one of the most widely determined parameters in managing organic pollution. The conventional BOD test includes a 5-day incubation period, so a more expeditious and reproducible method for assessment of this parameter is required. Trichosporon cutaneum, a microorganism formerly used in waste water treatment, has also been employed to construct a BOD biosensor. The dynamic system where the sensor was implemented consisted of a 0.1 M phosphate buffer at pH 7 saturated with dissolved oxygen which was transferred to a flow-cell at a rate of 1 mL/min. When the current reached a steady-state value, a sample was injected into the flow-cell at 0.2 mL/min. The steady-state current was found to be dependent on the BOD of the sample solution. After the sample was flushed from the flow-cell, the current of the microbial sensor gradually returned to its initial level. The response time of microbial sensors depends on the nature of the sample solution concerned. A linear relationship was foimd between the current difference (i.e. that between the initial and final steady-state currents) and the 5-day BOD assay of the standard solution up to 60 mg/L. The minimum measurable BOD was 3 mg/L. The current was reproducible within 6% of the relative error when a BOD of 40 mg/L was used over 10 experiments [128]. 

In principle, there are two possible ways to measure this effect. First, there is the end-point measurement (steady-state mode), where the difference is calculated between the initial current of the endogenous respiration and the resulting current of the altered respiration, which is influenced by the tested substances. Second, by kinetic measurement the decrease or the acceleration, respectively, of the respiration with time is calculated from the first derivative of the currenttime curve. The first procedure has been most frequently used in microbial sensors. These biosensors with a relatively high concentration of biomass have a longer response time than that of enzyme sensors. Response times of comparable magnitude to those of enzyme sensors are reached only with kinetically controlled sensors. 

The ability of the chemolithoautotrophic bacteria Thiobacillus ferrooxidans to oxidize Fe has already been utilized for construction of a microbial sensor for the determination of iron [101]. The limit of determination of this biosensor is 60 pmol 1" with a response time ranging from 30 s to 5 min, depending on the Fe +-concentration in the sample. 

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