Ordered mesoporous silica as supports for immobilization of biocatalyst

Mesoporous materials are of great interest to the materials community because of their potential applications for catalysis,separation of large molecules,medical implants,semiconductors,magnetoelectric devices.The thesis entitled 'Ordered Mesoporous Silica as supports for immobilization of Biocatalyst' presents how the pore size can be tuned without the loss in ordered structure for the entrapment of an industially important biocatalyst-amylase.Immobilization of enzymes on ordered mesoporous material has triggered new ooportunities for stabilizing enzymes with improved intrinsic and operational stabilities.

Development of Electrochemical Sensors for Various Pharmaceuticals

The development of electrochemical sensors is currently one of the active areas of research in analytical chemistry.Voltammetric sensors as an important class of electrochemical sensors are extensively used in pharmaceutical applications.In voltammetric analysis,many active compounds in dosage forms,in contrast to excipients,can be readily oxidised or reduced at the electrode surface by applying a potential.Chemically modified electrodes have great significance in the electrochemical determination of pharmaceuticals.The modification of electrode results in efficient determination of electroactive species at very lower potential without any major interferences.The present study involves the fabrication of 8 voltammetric sensors for the drugs Metronidazole Benzoate, Sulfamethoxazole, Acyclovir, Pam Chloride , Trimethoprim , Tamsulosin Hydrochloride and Ceftriaxone Sodium.Two sensors were developed for the drug tamsulosin hydrochloride while one sensor each was developed for the other drugs.

Studies on immobilization of Bacteria

Cell immnhilizatinn technology in a rapidly expanding arna in the endeavour of microbial fnrmentatiwn.During the lnmt 15 years anveral prnceafinn have been developed and more are in developmental atage of approaching commercial utilizatinn.In the present programme it was planned to develop an optimized process for the innobilization of alpha amylase producing Bacillus polymyxa (CBTB 25) an isolate obtained from Cochin University campus primarily for the production of alpha-amylase.Optimal concentration of support material that attributaa stability and maximal activity to the immobilized cell beads was determined using different concentrations of sodium aliginate as support and estimation of amylase production.An overeall assessment of the data obtained for the various studies conducted denotes that immobilized cells synthesize alpha-amylase at comparable rates with free cells and produce reducing sugara at a higher level than free cells.Results indicated that both phosphate and citrate buffers could be used for disrupting the immobilized beads since they enforced maximal release of cells through leaching from the beads within one hour.On comparative analysis it was observed that immobilized cells could synthesize alpha amylase at similar levels with free cells of B.polymyxa.On Co-immobilization of B.Polymyxa with S.cerevisiae,the co-immobilizate beads could effeciently convert starch directly to ethanol with a yield of 14.8% at 1 : 2 ratio.

Development of Macrocyclics based Electrochemical Sensors

Electroanalytical techniques represent a class of powerful and versatile analytical method which is based on the electrical properties of a solution of the analyte when it is made part of an electrochemical cell. They offer high sensitivity, accuracy, precision and a large linear dynamic range. The cost of instrumentation is relatively low compared to other instrumental methods of analysis. Many solid state electrochemical sensors have been commercialised nowadays. Potentiometry is a very simple electroanalytical technique with extraordinary analytical capabilities. Since valinomycin was introduced as an ionophore for K+, Ion Selective Electrodes have become one of the best studied and understood analytical devices. It can be used for the determination of substances ranging from simple inorganic ions to complex organic molecules. It is a very attractive option owing to the wide range of applications and ease of the use of the instruments employed. They also possess the advantages of short response time, high selectivity and very low detection limits. Moreover, analysis by these electrodes is non-destructive and adaptable to small sample volumes. It has become a standard technique for medical researchers, biologists, geologists and environmental specialists. This thesis presents the synthesis and characterisation of five ionophores. Based on these ionophores, nine potentiometric sensors are fabricated for the determination of ions such as Pb2+, Mn2+, Ni2+, Cu2+ and Sal- ion (Salicylate ion). The electrochemical characterisation and analytical application studies of the developed sensors are also described. The thesis is divided into eight chapters

Structural, electrical and electrochemical characterization of high frequency magnetron sputtered spinel oxide cathode films for lithium ion battery applications

The main challenges in the deposition of cathode materials in thin film form are the reproduction of stoichiometry close to the bulk material and attaining higher rates of deposition and excellent crystallinity at comparatively lower annealing temperatures. There are several methods available to develop stoichiometric thin film cathode materials including pulsed laser deposition; plasma enhanced chemical vapor deposition, electron beam evaporation, electrostatic spray deposition and RF magnetron sputtering. Among them the most versatile method is the sputtering technique, owing to its suitability for micro-fabricating the thin film batteries directly on chips in any shape or size, and on flexible substrates, with good capacity and cycle life. The main drawback of the conventional sputtering technique using RF frequency of 13.56MHz is its lower rate of deposition, compared to other deposition techniques A typical cathode layer for a thin film battery requires a thickness around one micron. To deposit such thick layers using convention RF sputtering, longer time of deposition is required, since the deposition rate is very low, which is typically 10-20 Å/min. This makes the conventional RF sputtering technique a less viable option for mass production in an economical way. There exists a host of theoretical and experimental evidences and results that higher excitation frequency can be efficiently used to deposit good quality films at higher deposition rates with glow discharge plasma. The effect of frequencies higher than the conventional one (13.56MHz) on the RF magnetron sputtering process has not been subjected to detailed investigations. Attempts have been made in the present work, to sputter deposit spinel oxide cathode films, using high frequency RF excitation source. Most importantly, the major challenge faced by the thin film battery based on the LiMn2O4 cathode material is the poor capacity retention during charge discharge cycling. The major causes for the capacity fading reported in LiMn2O4cathode materials are due to, Jahn-Teller distortion, Mn2+ dissolution into the electrolyte and oxygen loss in cathode material during cycling. The work discussed in this thesis is an attempt on overcoming the above said challenges and developing a high capacity thin film cathode material.

Immobilization of nitrifying bacterial consortia on wood particles for bioaugmenting nitrification in shrimp culture systems

Shrimp grow out systems under zero water exchange mode demand constant remediation of total ammonia nitrogen (TAN) andNO2 −–Nto protect the crop. To address this issue, aninexpensive and user-friendly technology using immobilized nitrifying bacterial consortia (NBC) as bioaugmentors has been developed and proposed for adoption in shrimp culture systems. Indigenous NBC stored at 4 °C were activated at room temperature (28 °C) and cultured in a 2 L bench top fermentor. The consortia, after enumeration by epifluorescence microscopy,were immobilized on delignifiedwood particles of a soft wood tree Ailantus altissima (300–1500 μm) having a surface area of 1.87m2 g−1. Selection of wood particle as substratumwas based on adsorption of NBC on to the particles, biofilm formation, and their subsequent nitrification potential. The immobilization could be achievedwithin 72 h with an initial cell density of 1×105 cells mL−1. On experimenting with the lowest dosage of 0.2 g (wet weight) immobilized NBC in 20 L seawater, a TAN removal rate of 2.4 mg L−1 within three days was observed. An NBC immobilization device could be developed for on site generation of the bioaugmentor preparation as per requirement. The product of immobilization never exhibited lag phase when transferred to fresh medium. The extent of nitrification in a simulated systemwas two times the rate observed in the control systems suggesting the efficacy in real life situations. The products of nitrification in all experiments were undetectable due to denitrifying potency, whichmade the NBC an ideal option for biological nitrogen removal. The immobilized NBC thus generated has been named TANOX (Total Ammonia Nitrogen Oxidizer)

Immobilization of Diastase α-amylase on to synthetic and natural polymers

Several natural and synthetic supports have been assessed for their efficiency for enzyme immobilization. Synthetic polymer materials are prepared by chemical polymerization using various monomers. As a kind of important carrier, synthetic polymer materials exhibit the advantages of good mechanical rigidity, high specific surface area, inertness to microbial attack, easy to change their surface characteristics, and their potential for bringing specific functional group according to actual needs. Hence, they have been widely investigated and used for enzyme immobilization. When it comes to the natural polymer materials, much attention has been paid to cellulose and other natural polymer materials owing to their wide range of sources, easy modification, nontoxic, and pollution-free, with a possibility of introducing wide variety of functional groups and good biocompatible properties. In this work report the use of synthetic polymer, polypyrrole and its derivatives and natural polymers coconut fiber and sugarcane bagasse as supports for Diastase α- amylase immobilization. An attempt was also made to functionalize both synthetic and natural polymers using Amino-propyl triethoxysilane. Supports and their immobilized forms were characterized via FT-IR, TG, SEM, XRD, BET and EDS techniques. Immobilization parameters were also optimized so as to prepare stable immobilized biocatalyst for starch hydrolysis.

Electrochemical studies of tamsulosin hydrochloride using multiwalled carbon nanotube-modified glassy carbon sensor

A differential pulse voltammetric sensor for the determination of tamsulosin hydrochloride (TAM) using multiwalled carbon nanotubes (MWNTs)–Nafion-modified glassy carbon electrode (GCE) has been developed. MWNTs were dispersed in water with the help of Nafion and were used to modify the surface of GCE via solvent evaporation. At MWNT-modified electrode, TAM gave a well-defined oxidation peak at a potential of 1084 mV in 0.1 M acetate buffer solution of pH 5. Compared to the bare electrode, the peak current of TAM showed a marked increase and the peak potential showed a negative deviation. The determination conditions, such as the amount of MWNT–Nafion suspension, pH of the supporting electrolyte and scan rate, were optimised. Under optimum conditions, the oxidation peak current was proportional to the concentration of TAM in the range 1 × 1023 M–3 × 1027 M with a detection limit of 9.8 × 1028 M. The developed sensor showed good stability, selectivity and was successfully used for the determination of TAM in pharmaceutical formulations and urine samples

Novel Electrochemical and Fluorescence Sensors for Food Additives and Neurotransmitters

There is an enormous demand for chemical sensors in many areas and disciplines including chemistry, biology, clinical analysis, environmental science. Chemical sensing refers to the continuous monitoring of the presence of chemical species and is a rapidly developing field of science and technology. They are analytical devices which transform chemical information generating from a reaction of the analyte into an measurable signal. Due to their high selectivity, sensitivity, fast response and low cost, electrochemical and fluorescent sensors have attracted great interest among the researchers in various fields. Development of four electrochemical sensors and three fluorescent sensors for food additives and neurotransmitters are presented in the thesis. Based on the excellent properties of multi walled carbon nanotube (MWCNT), poly (L-cysteine) and gold nanoparticles (AuNP) four voltammetric sensors were developed for various food additives like propyl gallate, allura red and sunset yellow. Nanosized fluorescent probes including gold nanoclusters (AuNCs) and CdS quantum dots (QDs) were used for the fluorescent sensing of butylated hydroxyanisole, dopamine and norepinephrine. A total of seven sensors including four electrochemical sensors and three fluorescence sensors have been developed for food additives and neurotransmitters.

Development of Electrochemical and Fluorescent Sensors

Chemical sensors have growing interest in the determination of food additives, which are creating toxicity and may cause serious health concern, drugs and metal ions. A chemical sensor can be defined as a device that transforms chemical information, ranging from the concentration of a specific sample component to total composition analysis, into an analytically useful signal. The chemical information may be generated from a chemical reaction of the analyte or from a physical property of the system investigated. Two main steps involved in the functioning of a chemical sensor are recognition and transduction. Chemical sensors employ specific transduction techniques to yield analyte information. The most widely used techniques employed in chemical sensors are optical absorption, luminescence, redox potential etc. According to the operating principle of the transducer, chemical sensors may be classified as electrochemical sensors, optical sensors, mass sensitive sensors, heat sensitive sensors etc. Electrochemical sensors are devices that transform the effect of the electrochemical interaction between analyte and electrode into a useful signal. They are very widespread as they use simple instrumentation, very good sensitivity with wide linear concentration ranges, rapid analysis time and simultaneous determination of several analytes. These include voltammetric, potentiometric and amperometric sensors. Fluorescence sensing of chemical and biochemical analytes is an active area of research. Any phenomenon that results in a change of fluorescence intensity, anisotropy or lifetime can be used for sensing. The fluorophores are mixed with the analyte solution and excited at its corresponding wavelength. The change in fluorescence intensity (enhancement or quenching) is directly related to the concentration of the analyte. Fluorescence quenching refers to any process that decreases the fluorescence intensity of a sample. A variety of molecular rearrangements, energy transfer, ground-state complex formation and collisional quenching. Generally, fluorescence quenching can occur by two different mechanisms, dynamic quenching and static quenching. The thesis presents the development of voltammetric and fluorescent sensors for the analysis of pharmaceuticals, food additives metal ions. The developed sensors were successfully applied for the determination of analytes in real samples. Chemical sensors have multidisciplinary applications. The development and application of voltammetric and optical sensors continue to be an exciting and expanding area of research in analytical chemistry. The synthesis of biocompatible fluorophores and their use in clinical analysis, and the development of disposable sensors for clinical analysis is still a challenging task. The ability to make sensitive and selective measurements and the requirement of less expensive equipment make electrochemical and fluorescence based sensors attractive.

Contributions of labile and resistant organic materials to the immobilization of inorganic soil N when used in the restoration of abandoned agricultural fields

We have examined the contributions sucrose and sawdust make to the net immobilization of inorganic soil N and assimilation of both C and N into microbial biomass when they are used as part of a restoration plan to promote the establishment of indigenous vegetation on abandoned agricultural fields on the Central Hungarian Plain. Both amendments led to net N immobilization. Sucrose addition also led to mobilization of N from the soil organic N pool and its immobilization into microbial biomass, whereas sawdust addition apparently immobilized soil N into a non-biomass compartment or a biomass component that was not detected by the conventional biomass N assay (CHCl3 fumigation and extraction). This suggests that the N was either cycled through the biomass, but not immobilized within it, or that it was immobilized in a protected biomass fraction different to the fraction into which N was immobilized in response to sucrose addition.

Promoting microbial immobilization of soil nitrogen during restoration of abandoned agricultural fields by organic additions

Application of organic materials to soils to enhance N immobilization into microbial biomass, thereby reducing inorganic N concentrations, was studied as a management option to accelerate the reestablishment of the native vegetation on abandoned arable fields on sandy soils the Kiskunsag National Park, Hungary. Sucrose and sawdust were used at three different topographic sites over 4 years. N availability and extractable inorganic N concentrations were significantly reduced in all sites. Soil microbial biomass C and microbial biomass N increased significantly following C additions, but the microbial C to microbial N ratio remained unaffected. It is concluded that the combined application of the rapidly utilized C source (sucrose) promoted N immobilization, whereas the addition of the slowly utilized C source (sawdust) maintained the elevated microbial biomass C and microbial biomass N in the field.

Contributions of labile and resistant organic materials to the immobilization of inorganic soil N when used in the restoration of abandoned agricultural fields

We have examined the contributions sucrose and sawdust make to the net immobilization of inorganic soil N and assimilation of both C and N into microbial biomass when they are used as part of a restoration plan to promote the establishment of indigenous vegetation on abandoned agricultural fields on the Central Hungarian Plain. Both amendments led to net N immobilization. Sucrose addition also led to mobilization of N from the soil organic N pool and its immobilization into microbial biomass, whereas sawdust addition apparently immobilized soil N into a non-biomass compartment or a biomass component that was not detected by the conventional biomass N assay (CHCl3 fumigation and extraction). This suggests that the N was either cycled through the biomass, but not immobilized within it, or that it was immobilized in a protected biomass fraction different to the fraction into which N was immobilized in response to sucrose addition.

Electrochemical determination of plant-derived leuco-indigo after chemical reduction by glucose

Electrochemical determination of redox active dye species is demonstrated in indigo samples contaminated with high levels of organic and inorganic impurities. The use of a hydrodynamic electrode system based on a vibrating probe (250 Hz, 200 mu m lateral amplitude) allows time-independent diffusion controlled signals to be enhanced and reliable concentration data to be obtained under steady state conditions at relatively fast scan rates up to 4 V s-1In this work the indigo content of a complex plant-derived indigo sample (dye content typically 30%) is determined after indigo is reduced by addition of glucose in aqueous 0.2 M NaOH. The soluble leuco-indigo is measured by its oxidation response at a vibrating electrode. The vibrating electrode, which consisted of a laterally vibrating 500 mu m diameter gold disc, is calibrated with Fe(CN)(6) 3-/4- in 0.1 M KCl and employed for indigo determination at 55, 65, and 75 C in 0.2 M NaOH. Determinations of the indigo content of 25 different samples of plant-derived indigo are compared with those obtained by conventional spectrophotometry. This comparison suggests a significant improvement by the electrochemical method, which appears to be less sensitive to impurities.

Electrochemical and sonoelectrochemical monitoring of indigo reduction by glucose

The reduction of indigo (dispersed in water) to leuco-indigo (dissolved in water) is an important industrial process and investigated here for the case of glucose as an environmentally benign reducing agent. In order to quantitatively follow the formation of leuco-indigo two approaches based on (i) rotating disk voltammetry and (ii) sonovoltammetry are developed. Leuco-indigo, once formed in alkaline solution, is readily monitored at a glassy carbon electrode in the mass transport limit employing hydrodynamic voltammetry. The presence of power ultrasound further improves the leuco-indigo determination due to additional agitation and homogenization effects. While inactive at room temperature, glucose readily reduces indigo in alkaline media at 65 degrees C. In the presence of excess glucose, a surface dissolution kinetics limited process is proposed following the rate law d eta(leuco-indigo)/dt = k x c(OH-) x S-indigo where eta(leuco-indigo) is the amount of leuco-indigo formed, k = 4.1 x 10(-9) m s(-1) (at 65 degrees C, assuming spherical particles of I gm diameter) is the heterogeneous dissolution rate constant,c(OH-) is the concentration of hydroxide, and Sindigo is the reactive surface area. The activation energy for this process in aqueous 0.2 M NaOH is E-A = 64 U mol(-1) consistent with a considerable temperature effects. The redox mediator 1,8-dihydroxyanthraquinone is shown to significantly enhance the reaction rate by catalysing the electron transfer between glucose and solid indigo particles. (c) 2006 Elsevier Ltd. All fights reserved.