Eric D. Conte

Microwave mediated distillation with solid-phase microextraction: determination of off-flavors, geosmin and methylisoborneol, in catfish tissue

Abstract Presented is a rapid distillation device for use with solid-phase microextraction (SPME). We apply this device specifically for determining two semivolatile off-flavor compounds, methylisoborneol and geosmin, in channel catfish. The presence of these two compounds in channel catfish filets results in unwelcome tastes. In the presented procedure, a catfish tissue sample is placed within a sample container located inside the microwave device. Microwave radiation is applied and distillates formed migrate through a condenser via a purge gas and are collected in a sample vial. A SPME fiber is placed within the stirred collected distillate and methylisoborneol and geosmin are extracted. Qualitative and quantitative results of these extractions are obtained using a gas chromatograph-ion trap mass spectrometer. This solventless technique results in detection limits far below the human threshold for these off-flavor compounds in channel catfish.

Effect of xanthan/enzyme-modified guar gum mixtures on the stability of whey protein isolate stabilized fish oil-in-water emulsions.

The effect of xanthan gum (XG) and enzyme-modified guar (EMG) gum mixtures on the physicochemical properties and oxidative stability of 2wt% whey protein isolate (WPI) stabilized oil-in-water (O/W) emulsions containing 20%v/v fish oil was investigated. EMG was obtained by hydrolyzing native guar gum using α-galactosidase enzyme. At higher gum concentrations (0.2 and 0.3wt%), the viscosity of the emulsions containing XG/EMG gum mixtures was significantly higher (P<0.05) of all emulsions. Increasing concentrations (0-0.3wt%) of XG/EMG gum mixtures did not affect the droplet size of emulsions. Microstructure images revealed decreased flocculation at higher concentrations. Primary and secondary lipid oxidation measurements indicated a slower rate of oxidation in emulsions containing XG/EMG gum mixtures, compared to XG, guar (GG), and XG/GG gum mixtures. These results indicate that XG/EMG gum mixtures can be used in O/W emulsions to increase physical and oxidative stabilities of polyunsaturated fatty acids in foods.

Solid-phase extraction sorbent consisting of alkyltrimethylammonium surfactants immobilized onto strong cation-exchange polystyrene resin.

Presented is a solid-phase extraction sorbent material composed of cationic alkyltrimethylammonium surfactants attached to a strong cation-exchange resin via ion-exchange. The original hydrophilic cation-exchange resin is made hydrophobic by covering the surface with alkyl chains from the hydrophobic portion of the surfactant. The sorbent material now has a better ability to extract hydrophobic molecules from aqueous samples. The entire stationary phase (alkyltrimethylammonium surfactant) is removed along with the analyte during the elution step. The elution step requires a mild elution solvent consisting of 0.25 M Mg2+ in a 50% 2-propanol solution. The main advantage of using a removable stationary phase is that traditionally utilized toxic elution solvents such as methylene chloride, which are necessary to efficiently release strongly hydrophobic species from SPE stationary phases, may now be avoided. Also, the final extract is directly compatible with reversed-phase liquid chromatography. The performance of this procedure is presented using pyrene as a test molecule.

Alkyltrimethylammonium surfactant-mediated extractions:: characterization of surfactant-rich and aqueous layers, and extraction performance

Most surfactants employed for extraction purposes contain strongly absorbing chromophores, and therefore cannot be used with the ultraviolet-visible HPLC detector because of the high background created. Alkyltrimethylammonium surfactants, which do not have strongly absorbing chromophores, have shown promise as an extractant compatible with HPLC-ultraviolet-visible detection. In our extraction procedure, alkyltrimethylammonium surfactants are added to a sample containing organic analytes in distilled water. Sodium chloride is next added, then the entire sample is shaken. Before centrifugation, 1-octanol is added to aid in the two phase formation of surfactant-rich and aqueous phases. In this paper, we present the results of our studies on the extraction behavior of an alkyltrimethylammonium surfactant technique using various organic compounds as test probes. Specifically studied are the extraction behavior of organic bases, isomers of varying polarity and a zwitterionic species that has different charges at various pH values. Results from multiple extractions to obtain quantitative recovery of analytes is also presented. The composition of each phase is elucidated through the interpretation of data obtained from thermogravimetric and carbon, hydrogen and nitrogen (CHN) instrumentation.

Efficient and inexpensive method for purification of heparin binding proteins.

Heparin binding (HB) proteins mediate a wide range of important cellular processes, which makes this class of proteins biopharmaceutically important. Engineering HB proteins may bring many advantages, but it necessitates cost effective and efficient purification methodologies compared to currently available methods. One of the most important classes of HB proteins are fibroblast growth factors (FGFs) and their receptors (FGFRs). In this study, we report an efficient off-column purification of FGF-1 from soluble fractions and purification of the D2 domain of FGFR from insoluble inclusion bodies, using a weak Amberlite cation (IRC) exchanger. FGF-1 and the D2 domain have been expressed in Escherichia coli and purified to homogeneity using IRC resin. This approach is an alternative to conventional affinity column chromatography, which exhibits several disadvantages, including time-consuming experimental procedures for purification and regeneration and results in the expensive production of recombinant proteins. Results of the heparin binding chromatography and steady state fluorescence experiments show that the FGF-1 and the D2 are in a native conformation. The findings of this study will not only aid an in-depth investigation of this class of proteins but will also provide avenues for inexpensive and efficient purification of other important biological macromolecules.

One-step synthesis of NaLu80 − xGdxF4:Yb183 +/Er23 +(Tm3 +) upconversion nanoparticles for in vitro cell imaging

Upconversion nanoparticles (UCNPs) possess a unique type of photoluminescence (PL) in which lower-energy excitation is converted into higher-energy emission via multi-photon absorption processes. In this work, we have used a facile one-step hydrothermal method promoted water solubility to synthesis NaLuGdF4:Yb3+/Er3+(Tm3+) UCNPs coated with malonic acid (MA). Scanning electron microscopy images and X-ray diffraction patterns reveal sphere-shaped UCNPs with an average size of ~80nm crystallized in the cubic NaLuF4 structure. The characteristic vibrations of cubic UCNPs have been taken into account by using Fourier-transform infrared spectroscopy. Based on PL studies, we have determined an optimal concentration of Gd3+ doping. The dependence of upconversion PL intensity on Gd3+ concentration is discussed via the results of magnetization measurements, which is related to the coupling/uncoupling of Gd3+ ions. Particularly, our study reveals that carboxyl-functionalized NaLuGdF4:Yb3+/Er3+(Tm3+) UCNPs have a relatively high cell viability with HeLa cells.