In vitro selection of DNA aptamers and their integration in a competitive voltammetric biosensor for azlocillin determination in waste water.

The uncontrolled usage of veterinary antibiotics has led to their widespread pollution in waterways and milk products. Potential impact of antibiotic residues on the environment and human health such as increased antibiotic resistance of microorganisms and triggering allergic reactions in humans have been reported. In this work, we developed a highly selective and sensitive voltammetric aptasensor for on-step, sensitive and low cost detection of azlocillin antibiotic, one of the broad spectrum ß-lactam antibiotics. The successful selection of DNA aptamers against azlocillin was accomplished using systemic evolution of ligands by exponential enrichment (SELEX) method. Fluorescence-binding assays showed dissociation constant of 55 nM for one of the selected aptamers (Az9). This aptamer was used to construct a competitive voltammetric aptasensor for azlocillin. A limit of detection of 1.2 pg/mL as well as remarkable selectivity against potential interfering agents, including amoxicillin, were achieved. This signal-off competitive sensor takes 30-50 min to complete the quantification of the target antibiotic. The sensor was challenged by detecting the target directly in complex environments such as tap and waste water where good recovery percentages were achieved.

Uni-molecular detection and quantification of selected ß-lactam antibiotics with a hybrid α-hemolysin protein pore.

Single-nanopores have recently been used to electrically detect a wide range of analytes. Similarly, using electrophysiology, we demonstrate how a system comprised of an ion channel formed by α-hemolysin (α-HL) and single-cyclic γ-cyclodextrin (γ-CD) molecule permits the detection of, and differentiation between three different antibiotics from the ß-lactam family. Specifically, histograms of the time between the successive binding events, and the residence time distributions of the antibiotic in the γ-CD molecular adapter vary with the antibiotic type. The results show that the association times of amoxicillin, azlocillin, and ampicillin are τ(on) = 2.1 ± 0.2, 2.2 ± 0.3, and 3.1 ± 0.4 ms, respectively. Interestingly, we found that the residence time of the bulkier and negatively charged azlocillin (τ(off) = 0.008 ± 0.0005 ms) is much less than that of ampicillin (τ(off) = 0.07 ± 0.005 ms) and amoxicillin (τ(off) = 0.1 ± 0.02 ms), even though the γ-CD-α-HL complex is anionic selective. The data were also used to estimate the standard free energy of binding between ampicillin to γ-CDs binding (-12 kJ mol(-1) [corrected]). The difference in association times might be due to γ-CDs-imposed steric hindrance or an energetically more expensive desolvation step for the antibiotics to gain access to the binding site in the CD. We suggest that this technique may be used to detect other analytes used in pharmaceutical applications.

Stability study of azlocillin sodium in glass bottles and PVC bags containing intravenous admixtures.

The kinetics of degradation of azlocillin sodium in four intravenous admixtures was investigated at different temperatures. The effect of temperature has been determined and from this data, by applying of Arrhenius-law, the stability of azlocillin sodium at 25 degrees C has been predicted and the t90 was determined. Admixtures containing azlocillin sodium (0.01 g ml-1) were prepared in 0.9% sodium chloride injection, in 5% dextrose solution, in 5% levulose solution and in Ringer's lactate solution. The admixtures were stored at 30 degrees, 40 degrees and 50 degrees C in either polyvinyl chloride bags and glass bottles. The change in the initial azlocillin sodium concentration was related to the type of intravenous solution. No dependence with material of container was found. After 24 hours, the change in the initial concentration of penicillin was less than 10% of the initial concentration in 0.9% sodium chloride and 5% levulose solution. However in Ringer's lactate and 5% glucose solution the t90 was lower. These results were found in agreement with experimental ones obtained at room temperature.

[Determination of azlocillin in the air]. / Opredelenie azlotsillina v vozdukhe.

A method for determination of air-borne azlocillin in production areas was developed. The method is based on redox reaction proceeding with reduction of the molybdenum blue heteropolycomplex. The reaction involves the antibiotic preliminarily hydrolyzed by alkali with heating. The blue solutions are photometered at 750 nm. The detection limit in the assay sample volume is 0.005 mg. The method may be useful in development of maximum permissible concentration of azlocillin and its control in ambient air of production areas.

Azlocillin levels in human tears and aqueous humor.

We administered 4 g of azlocillin sodium intravenously to 24 patients scheduled to undergo elective intraocular surgery and collected specimens of serum, tears, and aqueous humor from zero to six hours after infusion for assay by high-pressure liquid chromatography. We found that azlocillin is distributed into tears and penetrates into the aqueous humor of human volunteers with noninflamed eyes after a single intravenous dose. Levels of 4.17 micrograms/ml and 4.44 micrograms/ml were achieved in tears and aqueous humor, respectively.