Sol-Gel Films Doped with Enzymes and Banana Crude Extract as Sensing Materials for Spectrophotometric Determination.

Chromogenic enzymatic reactions are very convenient for the determination of various biochemically active compounds. Sol-gel films are a promising platform for biosensor development. The creation of sol-gel films with immobilized enzymes deserves attention as an effective way to create optical biosensors. In the present work, the conditions are selected to obtain sol-gel films doped with horseradish peroxidase (HRP), mushroom tyrosinase (MT) and crude banana extract (BE), inside the polystyrene spectrophotometric cuvettes. Two procedures are proposed: the use of tetraethoxysilane-phenyltriethoxysilane (TEOS-PhTEOS) mixture as precursor, as well as the use of silicon polyethylene glycol (SPG).In both types of films, the enzymatic activity of HRP, MT, and BE is preserved. Based on the kinetics study of enzymatic reactions catalyzed by sol-gel films doped with HRP, MT, and BE, we found that encapsulation in the TEOS-PhTEOS films affects the enzymatic activity to a lesser extent compared to encapsulation in SPG films. Immobilization affects BE significantly less than MT and HRP. The Michaelis constant for BE encapsulated in TEOS-PhTEOS films almost does not differ from the Michaelis constant for a non-immobilized BE. The proposed sol-gel films allow determining hydrogen peroxide in the range of 0.2-3.5 mM (HRP containing film in the presence of TMB), and caffeic acid in the ranges of 0.5-10.0 mM and 2.0-10.0 mM (MT- and BE-containing films, respectively). BE-containing films have been used to determine the total polyphenol content of coffee in caffeic acid equivalents; the results of the analysis are in good agreement with the results obtained using an independent method of determination. These films are highly stable and can be stored without the loss of activity for 2 months at +4 °C and 2 weeks at +25 °C.

Silica Xerogel Doped with Iron(III) as Sensor Material for Salicylhydroxamic Acid Determination in Urine.

Salicylhydroxamic acid (SHA) is used as antimicrobic medicine and its concentration has to be monitored in urine. For the first time, silica xerogels doped with iron(III) have been proposed as sensor materials for SHA determination in biological samples. Three xerogels with iron(III) content in the range of 0.04-1.74% wt have been synthesized. BET surface area of these xerogels has varied in the range of 696-529 m2/g and total pore volume has varied in the range of 0.92-0.23 cm3/g. Complex formation between immobilized iron(III) and salicylhydroxamic acid has been investigated with solid phase spectrophotometry and IR spectroscopy. Orange-brown iron(III)-SHA complex with 1:1 stoichiometry is formed at pH 1-4 with half-reaction time of 17 min. Silica xerogel doped with 0.33% wt iron(III)) has been used as sensor material for SHA solid phase spectrophotometric determination (LOD 1.4 mg/L (n = 3), analytical range 4-230 mg/L). Proposed sensor material has been applied for SHA determination in biological samples of synthetic and human urine. The proposed procedure is characterized by a good level of accuracy (recovery values 97-120%) and precision (RSD values 4-9%) and can be recommended for pharmacokinetic-pharmacodynamic studies of hydroxamic acid-based medications.

Silica and Silica-Titania Xerogels Doped with Iron(III) for Total Antioxidant Capacity Determination.

In order to design a sensor material for total antioxidant capacity determination we have prepared silica and silica-titania xerogels doped with iron(III) and modified with 1,10-phenanthroline. Titanium(IV) tetraethoxyde content in the precursors (titanium(IV) tetraethoxyde and tetraethyl orthosilicate) mixtures has been varied from 0 to 12.5% vol. Iron(III) concentrations in sol has been varied from 1 to 100 mM. The increase of titanium(IV) content has led to a decrease in BET surface area and average pore diameter and an increase of micropore surface area and volume, which has resulted in better iron(III) retention in the xerogels. Iron(III), immobilized in the xerogel matrix, retains its ability to form complexes with 1,10-phenanthroline and to be reduced to iron(II). Static capacities for 1,10-phenanthroline have been determined for all the iron(III) doped xerogels (0.207 mmol/g-0.239 mmol/g) and they are not dependent on the iron(III) content. Sensor materials-xerogels doped with iron(III) and modified with 1,10-phenanthroline-have been used for antioxidants (catechol, gallic and ascorbic acids, and sulphite) solid phase spectrophotometric determination. Limits of detection for catechol, gallic and ascorbic acids, and sulphite equal 7.8 × 10-6 M, 5.4 × 10-6 M, 1.2 × 10-5 M, and 3.1 × 10-4 M, respectively. The increase of titanium(IV) content in sensor material has led to an increase of the reaction rate and the sensitivity of determination. Proposed sensor materials have been applied for total antioxidant capacity (in gallic acid equivalents) determination in soft beverages, have demonstrated high stability, and can be stored up to 6 months at room temperature.

Agaricus bisporus Crude Extract: Characterization and Analytical Application.

In the present work crude Agaricus bisporus extract (ABE) has been prepared and characterized by its tyrosinase activity, protein composition and substrate specificity. The presence of mushroom tyrosinase (PPO3) in ABE has been confirmed using two-dimensional electrophoresis, followed by MALDI TOF/TOF MS-based analysis. GH27 alpha-glucosidases, GH47 alpha-mannosidases, GH20 hexosaminidases, and alkaline phosphatases have been also detected in ABE. ABE substrate specificity has been studied using 19 phenolic compounds: polyphenols (catechol, gallic, caffeic, chlorogenic, and ferulic acids, quercetin, rutin, dihydroquercetin, l-dihydroxyphenylalanine, resorcinol, propyl gallate) and monophenols (l-tyrosine, phenol, p-nitrophenol, o-nitrophenol, guaiacol, o-cresol, m-cresol, p-cresol). The comparison of ABE substrate specificity and affinity to the corresponding parameters of purified A. bisporus tyrosinase has revealed no major differences. The conditions for spectrophotometric determination have been chosen and the analytical procedures for determination of 1.4 × 10-4-1.0 × 10-3 M l-tyrosine, 3.1 × 10-6-1.0 × 10-4 M phenol, 5.4 × 10-5-1.0 × 10-3 M catechol, 8.5 × 10-5-1.0 × 10-3 M caffeic acid, 1.5 × 10-4-7.5 × 10-4 M chlorogenic acid, 6.8 × 10-5-1.0 × 10-3 M l-DOPA have been proposed. The procedures have been applied for the determination of l-tyrosine in food supplements, l-DOPA in synthetic serum, and phenol in waste water from the food manufacturing plant. Thus, we have demonstrated the possibility of using ABE as a substitute for tyrosinase in such analytical applications, as food supplements, medical and environmental analysis.

Determination of Food Oxalates Using Silica-Titania Xerogel Modified with Eriochrome Cyanine R.

The interaction of silica-titania xerogel with triphenylmethane dyes (pyrocatechol violet, chrome azurol S, eriochrome cyanine R) has been investigated to create a new sensor material for solid phase spectrophotometric determination of food oxalates. The complex forming reaction between xerogel incorporated titanium(IV) and triphenylmethane dyes has been studied; half-reaction periods, complex composition, equilibrium constants, and xerogel sorption capacity have been calculated for each dye. Eriochrome cyanine R (ECR) is characterized by the shortest half-reaction period, the smallest equilibrium constant, and the greatest capacity; it has been chosen for the sensor material construction because titanium(IV)-ECR complex is formed faster and can be destroyed easier than other studied complexes. The interaction of this sensor material with oxalates has been described: the presence of oxalates causes sensor material discoloration and the absorbance is used as analytical signal. The analytical range is 35-900 mg/L (LOD 10.5 mg/L, n = 7). High concentrations of interfering inorganic anions, organic acids, and sucrose did not affect oxalate determination. Proposed solid phase spectrophotometric procedure has been successfully applied for the determination of oxalates in food samples (sorrel, spinach, parsley, ginger, and black pepper) and the results are in good agreement with HPLC oxalate determination.

Silica-titania xerogel doped with Mo,P-heteropoly compounds for solid phase spectrophotometric determination of ascorbic acid in fruit juices, pharmaceuticals, and synthetic urine.

BACKGROUND: Ascorbic acid is one of the most important vitamins to monitor in dietary sources (juices and vitamins) and biological liquids. RESULTS: Silica and silica-titania xerogels doped with Mo,P-heteropoly compounds (HPC) have been synthesized varying titanium(IV) and HPC content in sol. Their surface area and porosity have been studied with nitrogen adsorption and scanning electron microscopy, their elemental composition has been studied with energy-dispersive X-ray analysis. The redox properties of the sensor material with sufficient porosity and maximal HPC content have been studied with potentiometry and solid phase spectrophotometry and it has been used for solid phase spectrophotometric determination of ascorbic acid. The proposed method is characterized by good selectivity, simple probe pretreatment and broad analytical range (2-200 mg/L, LOD 0.7 mg/L) and has been applied to the analysis of fruit juices, vitamin tablets, and synthetic urine. CONCLUSIONS: New sensor material has been used for simple and selective solid phase spectrophotometric procedure of ascorbic acid determination in fruit juices, vitamin tablets, and synthetic urine.Graphical abstractWe synthesized several silica-titania xerogels doped with Mo,P-heteropoly compounds, studied their properties, and designed the sensor material for solid phase spectrophotometric determination of ascorbic acid in fruit juices, pharmaceuticals, and synthetic urine.

Silica-titania xerogel for solid phase spectrophotometric determination of salicylate and its derivatives in biological liquids and pharmaceuticals.

BACKGROUND: Salicylic acid and its derivatives are widely used drugs with potential toxicity. The main areas of salicylate derivatives determination are biological liquids and pharmaceuticals analysis. RESULTS: Silica-titania xerogel has been used for solid phase spectrophotometric determination of various salicylate derivatives (salicylate, salicylamide, methylsalicylate). The reaction conditions influence on the interaction of salicylate derivatives with silica-titania xerogels has been investigated; the characteristics of titanium(IV)-salicylate derivatives complexes in solid phase have been described. The simple solid phase spectrophotometric procedures are based on the formation of xerogel incorporated titanium(IV) colored complexes with salicylate derivatives. A linear response has been observed in the following concentration ranges 0.1-5, 0.5-10 and 0.05-4.7 mM for salicylate, salicylamide, and methylsalicylate, respectively. The proposed procedures have been applied to the analysis of human urine, synthetic serum, and pharmaceuticals. CONCLUSIONS: The simple solid phase spectrophotometric procedures of salicylate derivatives determination based on the new sensor materials have been proposed for biological liquids and pharmaceuticals analysis. Graphical abstractComplexation of titanium (IV), incorporated in silica-titania xerogels (Si-Ti), with salicylate derivatives (L) resulting in yellow-colored xerogels (Si-Ti/Ln) has been proposed for salicylate derivatives determination in biological liquids and pharmaceuticals.

Silica and silica-titania sol-gel materials: synthesis and analytical application.

This review describes last decade progress in the synthesis of sol-gel materials with analytically relevant properties and their application for the determination of metal ions, non-metal and organic compounds. The following types of materials are discussed: silica sol-gel materials doped with various analytical reagents, organofunctional silica sol-gel materials, and silica titania sol-gel materials. The variety of obtained materials allows their application for a wide range of analytical systems: electrochemical sensors, solid phase spectrophotometrical and SIA determination, and also for test determination including the employment of "length-of-stain" indicator tubes. This review surveys the results of studies on mentioned above subjects and summarizes the works accomplished in the field by author's team in Moscow State University.