Selection and identification of a DNA aptamer for fluorescent detection of netilmicin.

Netilmicin (NET) is an antibiotic widely used in healthcare and agriculture, but it can accumulate in the environment to threat human health. Netilmicin (NET) is an antibiotic used for veterinary purposes, for human therapy and for agricultural purposes. Therefore, there is a need to develop high-sensitive measuring methods to detect NET. Aptamer-based detecting methods are highly sensitive, inexpensive, and portable. In this study, we developed an aptamer-based fluorescence method to detect and quantify NET. NET was first conjugated to magnetic beads by amidation reaction and then NET-coated beads were used as the stationary phase to isolate aptamers by systematic evolution of ligands by exponential enrichment (SELEX) screening method. After ten rounds of SELEX screening, 32 aptamers with NET-binding affinity were obtained and the candidate aptamer APT-21 was finally chosen by comprehensively comparing their secondary structure characters and NET-binding affinity. APT-21 bound to NET with high affinity (Kd = 194.1 nmol/L) and high specificity that it displayed low cross-binding activities on 7 different structural analogs. We also developed a fluorometric assay using SYBR Green I (SG-I) and the APT-21. Key experimental parameters were optimized, including buffer system, SG-I and APT-21 reaction time, SG-I concentration, and aptamer concentration, to improve the detecting sensitivity. Our results suggest that the low limit of detection (LOD) of this method reached a low level of 1.95 nM and it also exhibited a good linear range up to 200 nM. Moreover, we successfully applied our method to detect the NET spiked in tap water and river water with good recoveries in the range from 97% to 111%. In conclusion, our current study isolated a NET-specific aptamer and developed an aptamer-based quantification method, which is promising to apply to detect NET in environmental samples.

Colorimetric detection of ciprofloxacin in aqueous solution based on an unmodified aptamer and the aggregation of gold nanoparticles.

A colorimetric method is described for detection of the antibiotic ciprofloxacin (CIP) in aqueous solutions based on an unmodified CIP-aptamer and gold nanoparticles (AuNPs), which are regarded as a probe and an indicator, respectively. In the absence of CIP, aptamers can hybridize with poly dimethyl diallyl ammonium chloride (PDDA) to form a 'duplex' structure through electrostatic interactions, and AuNPs remain dispersed in solution, leading to a red solution and an obvious absorption peak at 520 nm. Contrarily, the CIP-aptamer can specifically bind to CIP after the introduction of CIP into the solution. Then PDDA is thus free to aggregate AuNPs. The solution turns blue from red accordingly, presenting a strong absorption at 650 nm. Hence, the concentration of CIP can be quantified through the changes of the absorption. This sensitive and selective colorimetric method for CIP detection has a good linear response (R = 0.9935) in the 20 to 300 nM CIP concentration range, with a limit of detection (LOD) of 0.215 nM. Importantly, the proposed aptasensor demonstrates great application potential in CIP determination in aqueous samples.

Adsorption characteristics of malic acid from aqueous solutions by weakly basic ion-exchange chromatography.

In this study, we reported the effects of temperature, malic acid loading concentration, and resin dose on malic acid adsorption by IRA-67 in batch experiments. The kinetic data well fitted the pseudo-second-order kinetic model. Both the equilibrium and ultimate adsorption slightly decreased with increased temperature from 303 to 323 K at 74.7 g/L malic acid loading concentration. The malic acid adsorption was revealed as a homogeneous adsorbent process by the Langmuir model and film diffusion process at loading concentrations of 18.2-94.5 g/L malic acid by the Boyd plot. The values of effective diffusion coefficient D(i) also increased with the temperature. Based on Eq. (15), the negative values of ΔG° and ΔH° revealed that the adsorption process was spontaneous and exothermic. The negative value of ΔS° also indicated the decrease in the solid-liquid interface randomness at this interface when malic acid is adsorbed by IRA-67.