Quantification Of Copper Ions Research Articles

Label-free multimodal analysis of copper ions at below permissible exposure limit in the aqueous medium

An anthraimidazoledione based amphiphilic dye molecule was synthesized that shows formation of tuneable charge-transfer state in solution, susceptible to change in pH, polarity and hydrogen bonding ability of the medium. The compound also showed formation of nanoscopic self-assembled structure in water medium. The probe molecule can achieve multimodal detection (colorimetric, fluorimetric and electrochemical) of copper ions as low as 0.3 ppm in the aqueous medium. Addition of copper leads to dose-dependent ratiometric change in solution color from yellow to purple. The mechanistic investigation indicates that the coordination of copper ions was possible via simultaneous engagement of both imidazole nitrogen ends and neighbouring hydroxyl unit. Not only optical property, the changes in microenvironment also influence the selectivity as well as sensitivity of the probe molecule towards Cu2+ ions. Further, the optical probe is used for detection as well as quantification of copper ions in natural water samples without any sample pretreatment. Low-cost, reusable paper strips are developed for rapid, on-location detection of residual Cu2+ in real-life samples.

Design of immunogens: The effect of bifunctional chelator on immunological response to chelated copper

To produce specific antibodies for the detection and quantification of copper ions, bifunctional chelators (BFCs) are commonly applied in the preparation of copper conjugates. However, some copper-chelator complexes exhibit limited stability under in vivo conditions. In this study, Cu2+ was coupled with carrier proteins via three different macrocyclic BFCs: p-SCN-Bn-DOTA, p-SCN-Bn-NOTA, and p-SCN-Bn-TETA. The stability in plasma and the immunogenicity of three copper immunoconjugates were compared. The chelators other than p-SCN-Bn-DOTA were very stable in plasma, with <9% dissociation of Cu2+ over 96 h. The immune response varied depending on the choice of chelator; notably, antisera from the Cu2+-NOTA-KLH conjugate demonstrated the best reactivity toward chelated Cu2+. p-SCN-Bn-NOTA, which showed significant advantages over the other chelators, was used for antibody production. The efficiency of immune-positive hybridoma production was satisfactory, and the resultant monoclonal antibodies (McAbs) 4B7 showed sensitivity (half-maximal inhibitory concentration (IC50) of 8.9 ng/mL) to chelated Cu2+, with a working range from 1.21 to 48.9 ng/mL. The recovery of Cu2+ from water samples was 85.7–108%, and the intra- and inter-assay coefficients of variation were 4.0–10.1% and 7.1–11.4%, respectively. Compared with previously reported McAb specific to Cu2+, DF4, the sensitivity of the newly developed assay was improved 100-fold. The results of this study indicate the utility of NOTA for the efficient generation of highly sensitive McAbs against Cu2+.

Fluorescence-based determination of the copper concentration in drinking water

Copper is a heavy metal, which is used in heat and electrical conductors and in a multitude of alloys in the technical context. Moreover, it is a trace element that is essential for the life of organisms but can cause toxic effects in elevated concentrations. Maximum limits in water and beverages exist. Here, the decrease of the fluorescence lifetime of green fluorescent protein (GFP) by Forster resonance energy transfer is used to measure the copper ion concentration in drinking water. Therefore, a system is developed that is based on a GFP sample in a predefined concentration. The GFP mutant can be excited with blue light. For binding of copper ions, a His-tag is included in the GFP. After measuring the fluorescence lifetime of pure GFP, the copper determination of the sample is performed by lifetime measurement. Therefore, the lifetime can be assigned to the copper concentration of the GFP-doped drinking water sample. In summary, a method for the quantification of copper ions based on changes of the fluorescence lifetime of GFP is developed, and the measurement of the copper concentration in water samples is performed.