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The speciation of Cu2+, Ni2+ and Zn2+ ions in the presence of the fluoroquinolones (FQs) moxifloxacin, ofloxacin, levofloxacin and ciprofloxacin, in human blood plasma was studied under physiological conditions by computer simulation. The speciation was calculated using an updated model of human blood plasma including over 6,000 species with the aid of the program Hyss2009. The identity and stability of metal-FQ complexes were determined by potentiometric (310 K, 0.15 mol/L NaCl), spectrophotometric, spectrofluorimetric, ESI-MS and 1H-NMR measurements. In the case of Cu2+ ion the concentration of main low molecular weight (LMW) plasma complex (Cu(Cis)His) is very slightly influenced by all examined FQs. FQs show much higher influence on main plasma Ni2+ and Zn2+ complexes: (Ni(His)2 and Zn(Cys)Cit, respectively. Levofloxacin exhibits the highest influence on the fraction of the main nickel complex, Ni(His)2, even at a concentration level of 3 × 10−5 mol/L. The same effect is seen on the main zinc complex, Zn(Cys)Cit. Calculated plasma mobilizing indexes indicate that ciprofloxacin possesses the highest mobilizing power from plasma proteins, toward copper ion, while levofloxacin is the most influential on nickel and zinc ions. The results obtained indicate that the drugs studied are safe in relation to mobilization of essential metal ions under physiological conditions. The observed effects were explained in terms of competitive equilibrium reactions between the FQs and the main LMW complexes of the metal ions.

In this work a new enzymatic method for the determination of uric acid in human serum has been developed. The method is based on the oxidative coupling reaction between the N-methyl-N-(4-aminophenyl)-3-methoxyaniline (NCP) reagent and the hydrogen donor reagent, N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methylaniline (TOOS), in the system involving three enzymes: uricase, peroxidase and ascorbate oxidase. Using this method uric acid could be determined in concentrations up to 1.428 mmol/L, with a relative standard deviation of up to 1.8 %. The effect of the medium pH and the NCP concentration on the linearity of the chromogen absorbance versus the uric acid concentration curve was investigated. The influence of the uricase activity on the maximum rate of uric acid oxidation was also examined. The use of the NCP reagent demonstrated a more precise and more sensitive determination of the uric acid compared to the determination with 4-aminoantipyrine (4-AA) as the coupling regent. The sensitivity of the method determined from the calibration curve was 0.71 absorbance units per mmol/L of uric acid; the limit of detection was LOD = 0.0035 mmol/L and the limit of quantification was LOQ = 0.015 mmol/L of uric acid.

A sensitive and specific enzymatic method has been developed for the determination of the total and free cholesterol in human serum based on the application of the newly-synthesized N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline, (DAOS) Trinder's reagent. Using the proposed method, cholesterol could be determined in the concentration range of 0-15 mmol/L with a relative standard deviation of up to 1.1%. In order to find the optimal experimental conditions for the application of the DAOS reagent, the influence of its concentration on the linearity of the method, the influence of pH, as well as the influence of the activities of cholesterolesterase and cholesteroloxidase were examined. The obtained results of the determination of the total and free cholesterol were compared to the results obtained by the application of the most frequently employed enzymatic method in clinical practice, based on phenol as a reagent. The sensitivity of the method was 0.070 A/mmol/L, the limit of detection DL=0.03 mmol/L and the limit of quantification QL=0.09 mmol/L of cholesterol.

Abstract: An isocratic RP‐HPLC method for the separation and identification of selected flavonoids (quercetin, rutin, luteolin‐7‐O‐glucoside, kaempferol and kaempferol‐3‐O‐glucoside) in commercial berry juices (blackcurrant, blueberry, red raspberry and cherry) was developed with the aid of central composite design and response surface methodology. The optimal separation conditions were a mobile phase of 85:15 (% v/v) water–acetonitrile, pH 2.8 (adjusted with formic acid), flow rate 0.5 mL min−1 and column temperature 35°C. The obtained levels of bioflavonoids (mg per 100 mL of juice) were as follows: for quercetin, ca. 0.21–5.12; for kaempferol, ca. 0.05–1.2; for rutin, ca. 0.4–6.5; for luteolin‐7‐O‐glucoside, ca. 5.6–10.2; and for kaempferol‐3‐O‐glucoside, ca. 0.02–0.12. These are considerably lower than the values in fresh fruits. Total phenolic, flavonoid and anthocyanin contents were determined spectrophotometrically. Total flavonoid content varied as follows: blackcurrant > blueberry > red raspberry > cherry. The antioxidant activity of juice extracts (DPPH and ABTS methods) expressed as IC50 values varied from 8.56 to 14.05 mg L−1. These values are ~2.5–3 times lower than quercetin, ascorbic acid and Trolox®, but compared with rutin and butylhydroxytoluene, berries show similar or better antioxidant activity by both the DPPH and ABTS methods. [ABSTRACT FROM AUTHOR]

The main purpose of this study was to isolate airborne fungi and assess seasonal variations in air contamination with their particulates by determining the levels of their propagules in the nave and exonarthex of a church. We also monitored indoor microclimate as a determining factor for fungal proliferation on wall paintings, spore release, and transmission through the air. The temperature and relative humidity of the nave favoured fungal growth. A total of 33 fungi were isolated, mainly of the phylum Ascomycota, and to the lesser extent of the phyla Zygomycota and Basidiomycota. The most common were the fungi of the genera Penicillium and Aspergillus (23.55 % and 20.58 %, respectively). Sørensen's quotient of similarity (0.37) suggests moderate species overlap and constant exchange of fungal propagules between the nave and exonarthex. The autumn had the highest diversity, with 17 documented taxa, followed by the summer and the winter. The spring had only eight taxa. Quantitative analysis of the airborne mycobiota in the nave (430±84.85 to 1880±106.07 CFU m-3) and exonarthex (715±59.62 to 2295±91.92 CFU m-3) showed very high contamination throughout the year, with values exceeding the maximum permissible concentrations by most standards. Many of the fungi determined in this study are known for their biodeteriogenic, toxigenic, and allergenic properties, and are a threat not only to occasional visitors and staff, but also to valuable works of art decorating nave walls.