Sequential injection method for bi-parametric determination of iron and manganese in soil leachates.

The aim of this work was to develop a sequential injection (SI) method for the determination of the micronutrients iron and manganese, in soil leachates, as a tool to assess potential groundwater contamination. The described sequential injection method was based on the reaction of iron with chelator MRB12, a greener alternative chromogenic reagent, and the reaction of manganese with zincon, within a single manifold. The developed SI method enabled the determination of iron in the range 0.10-1.00 mg L-1, and manganese in the range 0.25-2.5 mg L-1 with a limit of detection of 0.08 mg L-1 for iron and 0.24 mg L-1 for manganese. The determination of both parameters was made in 6 minutes, in triplicate. The application to monitor laboratory scale soil core columns (LSSCs), as a simulation of the soil leaching process, proved its efficiency to assess potential contamination of ground waters. Iron and manganese contents were effectively analysed in two different scenarios to mimic the leaching process with rainwater and fertilizer.

Foliar application of 3-hydroxy-4-pyridinone Fe-chelate [Fe(mpp)3 ] induces responses at the root level amending iron deficiency chlorosis in soybean.

Iron (Fe) deficiency chlorosis (IDC) affects the growth of several crops, especially when growing in alkaline soils. The application of synthetic Fe-chelates is one of the most commonly used strategies in IDC amendment, despite their associated negative environmental impacts. In a previous work, the Fe-chelate tris(3-hydroxy-1-(H)-2-methyl-4-pyridinonate) iron(III) [Fe(mpp)3 ] has shown great potential for alleviating IDC in soybean (Glycine max) in the early stages of plant development under hydroponic conditions. Herein, its efficacy was verified under soil conditions in soybean grown from seed to full maturity. Chlorophyll levels, plant growth, root and shoot mineral accumulation (K, Mg, Ca, Na, P, Mn, Zn, Ni, and Co) and FERRITIN expression were accessed at V5 phenological stage. Compared to a commonly used Fe chelate, FeEDDHA, supplementation with [Fe(mpp)3 ] led to a 29% higher relative chlorophyll content, 32% higher root biomass, 36% higher trifoliate Fe concentration, and a twofold increase in leaf FERRITIN gene expression. [Fe(mpp)3 ] supplementation also resulted in increased accumulation of P, K, Zn, and Co. At full maturity, the remaining plants were harvested and [Fe(mpp)3 ] application led to a 32% seed yield increase when compared to FeEDDHA. This is the first report on the use of [Fe(mpp)3 ] under alkaline soil conditions for IDC correction, and we show that its foliar application has a longer-lasting effect than FeEDDHA, induces efficient root responses, and promotes the uptake of other nutrients.

Design and Characterization of Sodium Alginate and Poly(vinyl) Alcohol Hydrogels for Enhanced Skin Delivery of Quercetin.

Nature has led to the discovery of biopolymers with noteworthy pharmaceutical applications. Blended biopolymers have demonstrated promising characteristics when compared with their individual counterparts. Sodium alginate (SA) is a marine polymer that has demonstrated the ability to form hydrogels, an interesting property for the development of cutaneous formulations. Predicting the good performance of blended biopolymers, a novel series of hybrid hydrogels based on SA and poly(vinyl) alcohol (PVA) were prepared. Quercetin, a natural polyphenolic flavonoid commonly found in fruits and vegetables, is widely known for its strong anti-inflammatory and antioxidant activity, thus with potential applications against melanoma, dermatitis, psoriasis, and skin ageing. Here, hydrogels were produced at different ratios of SA and PVA. The surface morphology, structure, interaction of polymers, the capacity to absorb water and the entrapment efficiency of quercetin were evaluated for the blended hydrogels. Targeting the cutaneous application of the formulations, the rheological properties of all unloaded and quercetin-loaded hydrogels revealed pseudoplastic behavior, evidence of non-thixotropy, good resistance to deformation, and profile maintenance with temperatures ranging from 20 °C up to 40 °C. The incorporation of quercetin in the hydrogel retained its antioxidant activity, confirmed by radical scavenging assays (ABTS and DPPH). The permeability of quercetin through the skin showed different penetration/permeation profiles according to the hydrogel's blend. This behavior will allow the selection of SA-PVA at 2/1 ratio for a local and prolonged skin effect, making the use of these hydrogels a good solution to consider for the treatment of skin ageing and inflammation.

Greener and wide applicability range flow-based spectrophotometric method for iron determination in fresh and marine water.

A flow-based method for the spectrophotometric determination of iron in recreational waters, both fresh and marine (variable salinity content), was developed. For that purpose, 3-hydroxy-4-pyrydinone ligand functionalized with an ether function was synthetized and used as chromogenic chelator (1-(3'-methoxypropyl)-2-methyl-3-benzyloxy-4-(1H)pyridinone - MRB13) for iron quantification. This water-soluble reagent was previously reported as a greener alternative to quantify iron, due to its low toxicity and a more environmental friendly synthesis. Furthermore, it also displayed a high affinity and specificity for iron. With the main objective of quantifying iron in a variety of water types (different matrices and iron content), two strategies were developed, one of them including on-line solid-phase extraction (SPE), and the other without resorting to a SPE process. Water matrix clean-up and iron enrichment was achieved using a nitrilotriacetic acid resin column. The potential interference of metal ions usually present in water samples was assessed and no significant interference (<10%) was observed. The limits of detection were 11 and 2.9 µg L-1 without and with SPE, respectively. For one determination (three replicates), the corresponding consumption of MRB13 is 90 µg, sodium hydroxide is 1.4 mg, and boric acid is 5.6 mg. The method was applied to certified water samples and the results were in agreement with certified values. The developed method was also applied to fresh and marine water, and recovery ratios of 103 ± 4 and 101 ± 7 without and with SPE, respectively, were achieved.

Use of an ether-derived 3-hydroxy-4-pyridinone chelator as a new chromogenic reagent in the development of a microfluidic paper-based analytical device for Fe(III) determination in natural waters.

This article reports on the development and validation of a disposable microfluidic paper-based analytical device (µPAD) for on-hand, in-situ, and cheap Fe(III) determination in natural waters complying with World Health Organization guidelines. The developed µPAD used 3-hydroxy-4-pyridinone (3,4-HPO) as a colour reagent due to its considerably lower toxicity than traditionally used iron analytical reagents. It was selected among a group of hydrophilic 3,4-HPO chelators containing ether-derived chains in their structure which were prepared using green methods. The relatively high water solubility of these chelators improved the detection limit and applicability as µPAD reagents. Under optimal conditions, the µPAD is characterised by a quantification range between 0.25 and 2.0 mg/L, a detection limit of 55 µg/L and 15 min of analysis time. The signal stability extends up to 4 h and the device is stable for at least one month. The reagent consumption is below 0.2 mg per analysis and the µPAD method was validated by analysis certified reference materials and by comparison with atomic absorption results (RD < 10%). The newly developed µPAD was successfully applied to the determination of iron in river, well and tap waters with no need of any prior sample pre-treatment.

Insights on the relationship between structure vs. toxicological activity of antibacterial rhodamine-labelled 3-hydroxy-4-pyridinone iron(III) chelators in HepG2 cells.

In the present study we investigated the in vitro hepatotoxicity of a set of rhodamine-labelled 3-hydroxy-4-pyridinones (3,4-HPO) that had previously demonstrated significant inhibitory effect in the intramacrophagic growth of Mycobacterium avium. Our aim was to establish a correspondence between the molecular structure and the in vitro toxicological activity of these compounds. The impact of a set of bidentate (MRB2, MRB7, MRB8, and MRB9) and hexadentate (MRH7, MRH8, and MRH10) chelators on cellular metabolic competence and membrane integrity was investigated in HepG2 cells. Our findings indicate that: a) hexadentate chelators are more cytotoxic than parent bidentate ligands; b) disruption of cell membrane and metabolic competence only occurred after 5 days, at the highest concentrations tested; c) strict correlation between bacteriostatic activity and in vitro toxicity was observed, which seems to be directly dependent on the size of the molecule and on the hydrophilic/lipophilic balance; d) among the set of bidentate ligands, carboxyrhodamine derivatives (amide linker) presented lower detrimental effects, when compared with rhodamine B isothiocyanate chelators (thiourea linker); e) contrarily, for the hexadentate series, rhodamine B isothiocyanate derivatives are less cytotoxic to HepG2 cells than carboxyrhodamine molecules; and f) for all compounds tested, when the substituents of the nitrogen atom were switched from ethyl to methyl, an increment of toxicity was observed. Overall, all chelators seem to display suitable in vitro toxicological potential to combat fast grow bacteria. According to their in vitro pharmacological: toxicological potential ratio, MRH7 and MRH8 may be considered as the most suitable compounds to undergo further pre-clinical development studies.

Microsequential injection lab-on-valve system for the spectrophotometric bi-parametric determination of iron and copper in natural waters.

The determination of iron and copper exploiting a microsequential injection lab-on-valve system with online spectrophotometric detection is described. A new, environmental friendly 3-hydroxy-4-pyridinone chelator, functionalized with a polyethylene glycol chain (MRB12) to improve water solubility, was used for iron determination. For copper determination, 1-(2-pyridylazo)-2-naphthol (PAN) was used. Different parameters affecting the formation of the complexes were studied, namely sample volume, reagent concentration, and buffer composition and concentration. The optimized conditions, 150µL of sample volume and 250mgL-1 of MRB12 for iron determination, and 200µL of sample and 120mgL-1 of PAN for copper determination, enabled an LOD of 15 and 18µgL-1 for iron and copper, respectively. The robustness of the developed procedure was assessed by the calculation of the relative standard deviation (RSD), 5% for iron determination and 2% for copper determination. The accuracy of the method was assessed comparing the results with two certified samples (RD<7.5%) and calculating recovery percentages with five river water samples (average<107%).

Effect of tris(3-hydroxy-4-pyridinonate) iron(III) complexes on iron uptake and storage in soybean (Glycine max L.).

Iron deficiency chlorosis (IDC) is a serious environmental problem affecting the growth of several crops in the world. The application of synthetic Fe(III) chelates is still one of the most common measures to correct IDC and the search for more effective Fe chelates remains an important issue. Herein, we propose a tris(3-hydroxy-4-pyridinonate) iron(III) complex, Fe(mpp)3, as an IDC corrector. Different morphological, biochemical and molecular parameters were assessed as a first step towards understanding its mode of action, compared with that of the commercial fertilizer FeEDDHA. Plants treated with the pyridinone iron(III) complexes were significantly greener and had increased biomass. The total Fe content was measured using ICP-OES and plants treated with pyridinone complexes accumulated about 50% more Fe than those treated with the commercial chelate. In particular, plants supplied with compound Fe(mpp)3 were able to translocate iron from the roots to the shoots and did not elicit the expression of the Fe-stress related genes FRO2 and IRT1. These results suggest that 3,4-HPO iron(III) chelates could be a potential new class of plant fertilizing agents.

Relevant interactions of antimicrobial iron chelators and membrane models revealed by nuclear magnetic resonance and molecular dynamics simulations.

The dynamics and interaction of 3-hydroxy-4-pyridinone fluorescent iron chelators, exhibiting antimicrobial properties, with biological membranes were evaluated through NMR and molecular dynamics simulations. Both NMR and MD simulation results support a strong interaction of the chelators with the lipid bilayers that seems to be strengthened for the rhodamine containing compounds, in particular for compounds that include ethyl groups and a thiourea link. For the latter type of compounds the interaction reaches the hydrophobic core of the lipid bilayer. The molecular docking and MD simulations performed for the potential interaction of the chelators with DC-SIGN receptors provide valuable information regarding the cellular uptake of these compounds since the results show that the fluorophore fragment of the molecular framework is essential for an efficient binding. Putting together our previous and present results, we put forward the hypothesis that all the studied fluorescent chelators have access to the cell, their uptake occurs through different pathways and their permeation properties correlate with a better access to the cell and its compartments and, consequently, with the chelators antimicrobial properties.

Rhodamine labeling of 3-hydroxy-4-pyridinone iron chelators is an important contribution to target Mycobacterium avium infection.

We have recently demonstrated that tripodal hexadentate chelators, based on 3-hydroxy-4-pyridinone units, can limit the access of iron to bacteria and have a significant inhibitory effect in the intramacrophagic growth of Mycobacterium avium. The results showed that the chelation of iron is a determinant although not sufficient property for antimicrobial activity. The rhodamine B isothiocyanate labelled chelator (MRH7) exhibited the strongest inhibitory activity and was identified as a lead compound since a dose response effect was observed. Significant inhibition of M. avium growth was achieved at a concentration as low as 1 µM. To identify key molecular features essential for the biological activity we designed parent hexadentate and bidentate chelators, in which different structural groups are introduced in the molecular framework. Herein, we report the work concerning three novel fluorescent chelators: a hexadentate ligand labelled with 5(6)-carboxytetramethylrhodamine (MRH8) and two 3-hydroxy-4-pyridinone fluorescent bidentate ligands labelled with rhodamine B isothiocyanate (MRB7) and 5(6)-carboxytetramethylrhodamine (MRB8). The results show that all fluorescent chelators are capable of restricting the intramacrophagic growth of M. avium and that the inhibitory effect is dependent on the fluorophore. In fact, for compounds bearing the same fluorophore the results obtained with the hexadentate or bidentate chelator (MRH7/MRB7 or MRH8/MRB8) are identical as long as the appropriate stoichiometric amount of chelator is used. The inhibitory effect of the rhodamine B isothiocyanate labelled compounds (MRH7 and MRB7) is significantly greater than that observed for the other two chelators, thus pointing out the significance of the rhodamine B isothiocyanate molecular fragment.