The determination of the solubility of organic solvents in mineral acids

A knowledge of the solubility of organic solvents in aqueous solutions of mineral acids is shown to be important in solvent extraction studies. The method described depends upon the direct measurement of the amount of solvent thrown out of the aqueous phase on neutralization. The results of experiments on the solubility of iso-propyl ether in aqueous hydrochloric acid are given. The accuracy, limitations, and uses of the method are discussed.     

Evaporative micro-particle self assembly influenced by capillary evacuation

As evaporation does not incur energy introduction, the droplet coffee-stain patterning approach is attractive for biochemical tests conducted in the field or in third world environments. A practical strategy uses chemically functionalized microbeads for the coffee stain deposition process. From an application perspective, it will be necessary to minimize the coffee stain deposition time, as evaporation, depending on the volume of the droplet, can be a slow process. The introduction of a porous media will generate a capillary flow (or wicking) that removes any remnant liquid in the droplet, thus permitting it to be done inexpensively and in the field. Using optical profilometry, we were able to establish that polystyrene microspheres developed more copious and defined single ring coffee depositions than silica of the same size and concentration in a suspension. In analyzing the droplet capillary evacuation process with a porous media, we found the liquid bridge formed during the later stages to rupture and leave behind some liquid material for a second stage evaporation process. This was responsible for a two ring structure that was more visible with silica microspheres. A high degree of hysteresis of the contact angle was found to develop at the contact line in which values below 5° could be achieved. Dynamic observations showed the copious and dense packing of polystyrene particles to be more resistant to ring break up from the evacuation flow. Nevertheless, erosion of the back array portions of the ring was evident notwithstanding either type of microsphere used.     

Surface tension drawing of liquid from microplate capillary wells

Pressure differentials are routinely used to actuate flow in capillaries. We advance here an alternative means of flow generation that capitalizes on the extension of a liquid bridge achieved by the drawing of a rod through the action of surface tension. This meets the exigencies of creating controllable flow using simpler and more compact means. We found the ability to generate controllable flow to be strongly affected by the liquid bridge sustaining features, and that the use of rod diameters larger than the capillary was more conducive. The extensional flow resulting from the rupture of the liquid bridge was also found to have a strong circulation component which facilitated mixing. The approach here is highly amenable for use in capillary well microplates which have significant advantages over standard microplates. The features of this approach offer usage possibilities in biochemical applications in the field, such as in the leukocyte cell adhesion and hemagglutination tests of blood samples.     

Facile, divergent route to bis-Zn(II)dipicolylamine type chemosensors for pyrophosphate

A new, two-step synthesis has been developed for a series of bis-DPA-type ligands whose dinuclear Zn(II) complexes function as fluorescent anion sensors. The Zn(II) complexes exhibit good selectivity for PP_i over other anions in aqueous medium (pH 7.5) and may be used to monitor the extent of enzyme-catalysed reactions, in which PP_i is produced or consumed.     

A filter paper-based assay for laboratory evolution of hydrolases and dehydrogenases

Industrially important enzyme classes such as hydrolases and dehydrogenases are often not amenable to laboratory evolution methods due to a lack of sensitive and reliable high-throughput screening (HTS) systems. We developed a conceptually novel and technically simple high-throughput screening system based on detection of volatile aldehydes with the sensitive reagent Purpald (4-amino-3-hydrazino-5-mercapto-1,2,4-triazole). The aldehyde detection takes place on a filter-paper that is pre-soaked with Purpald and covers the microtiter plate. The filter paper-based Purpald assay separates aldehyde detection from biocatalytical conversion and thereby avoids interferences from biological materials with assay components. This screening principle allows, to our knowledge, for the first time to determine the synthetic activity of hydrolases such as lipases and esterases in organic solvents in a 96-well whole-cell format. Its simplicity and cost-effectiveness make the reported HTS system suitable as fast pre-screen in laboratory evolution experiments and for semi-quantitative assays of improved mutants.     

The design and synthesis of inhibitors of pantothenate synthetase

Pantothenate synthetase catalyses the ATP-dependent condensation of D-pantoate and beta-alanine to form pantothenate. Ten analogues of the reaction intermediate pantoyl adenylate, in which the phosphodiester is replaced by either an ester or sulfamoyl group, were designed as potential inhibitors of the enzyme. The esters were all modest competitive inhibitors, the sulfamoyls were more potent, consistent with their closer structural similarity to the pantoyl adenylate intermediate.     

Fluorescent Zn chemosensors, functional in aqueous solution under environmentally relevant conditions

The synthesis and evaluation of two new ratiometric chemosensors for the quantification of potentially toxic free Zn²⁺ ions in aqueous solutions are described. Both sensors show high selectivity for Zn²⁺ over other cations, and are functional at environmentally relevant pH with detection limits of 0.05 μM for free Zn²⁺.     

Effect of an encapsulated bubble in inhibiting droplet sliding

The transport of liquid droplets on surfaces carrying reactants offers advantages in the creation of fluidic devices crucial for life science applications. In a majority of situations, a selection of these droplets on a surface, rather than all of them, will need to be moved at any one time. It is a formidable challenge to deliver the motive energy source only to specific droplets while leaving the others unmoved. Here, we describe an alternative novel solution of momentarily pinning specific droplets to the surface while allowing the rest to be moved. We demonstrate this concept via the injection of a sizable bubble that is attached to a PTFE surface within a droplet. This then affects the contact line of the droplet, pinning it despite the introduction of an incline that will normally result in sliding. The use of bubbles offers easy release of pinning at will by simple rupture using mechanical means.