Rapid determination of free anthocyanins in foodstuffs using high performance liquid chromatography

Anthocyanins constitute a major group of natural pigments, and they are responsible for the colours of fruits and vegetables. A rapid and feasible assay procedure for the determination of free forms of the six most abundant anthocyanins in foods is described. The 3-glucoside forms of pelargonidin, cyanidin, peonidin, delphinidin, petunidin and malvidin with the aglycone cyanidin (as internal standard) were separated by gradient elution and quantified using HPLC-DAD within 18 min. A fast sample preparation step was employed which allows direct injection of samples to the chromatograph without need of chemical extraction. Testing on 28 different vegetable, fruit and processed commercial product samples demonstrated applicability in the concentration range of about 80–420 ng/mL with an accuracy of 99.2 ± 0.2% and an average precision of 0.8%. The method was suggested as a cheap and robust alternative to the previous ones that employ multi-step sample treatment protocols.     

Nonporous monosize polymeric sorbents: Dye and metal chelate affinity separation of lysozyme

Lysozyme adsorption onto dye‐attached nonporous monosize poly(2‐hydroxyethyl‐methacrylate‐methylmethacrylate) [poly(HEMA‐MMA)] microspheres was investigated. Poly(HEMA‐MMA) microspheres were prepared by dispersion polymerization. The monochloro‐triazine dye, Cibacron Blue F3GA, was immobilized covalently as dye–ligand. These dye‐affinity microspheres were used in the lysozyme adsorption–desorption studies. The effect of initial concentration of lysozyme and medium pH on the adsorption efficiency of dye‐attached and metal‐chelated microspheres were studied in a batch reactor. Effect of Cu(II) chelation on lysozyme adsorption was also studied. The nonspecific adsorption of lysozyme on the poly(HEMA‐MMA) microspheres was 3.6 mg/g. Cibacron Blue F3GA attachment significantly increased the lysozyme adsorption up to 247.8 mg/g. Lysozyme adsorption capacity of the Cu(II) incorporated microspheres (318.9 mg/g) was greater than that of the Cibacron Blue F3GA‐attached microspheres. Significant amount of the adsorbed lysozyme (up to 97%) was desorbed in 1 h in the desorption medium containing 1.0M NaSCN at pH 8.0 and 25 mM EDTA at pH 4.9. In order to examine the effects of separation conditions on possible conformational changes of lysozyme structure, fluorescence spectrophotometry was employed. We conclude that dye‐ and metal‐chelate affinity chromatography with poly(HEMA‐MMA) microspheres can be applied for lysozyme separation without causing any significant changes and denaturation. Repeated adsorption/desorption processes showed that these novel dye‐attached monosize microspheres are suitable for lysozyme adsorption. © 2000 John Wiley & Sons, Inc. J Appl Polym Sci 76: 115–124, 2000     

New metal chelate sorbent for albumin adsorption: Cibacron Blue F3GA-Zn(II) attached microporous poly(HEMA) membranes

Poly(2-hydroxyethyl methacrylate) [poly(HEMA)] membranes were prepared by UV-initiated photopolymerization of HEMA in the presence of an initiator (α-α′-azobis-isobutyronitrile, AIBN). The triazine dye Cibacron Blue F3GA was attached as an affinity ligand to poly(HEMA) membranes, covalently. These affinity membranes with a swelling ratio of 58% and containing 10.7 mmol Cibacron Blue F3GA/m² were used in the albumin adsorption studies. After dye-attachment, Zn(II) ions were chelated within the membranes via attached-dye molecules. Different amounts of Zn(II) ions [650–1440 mg Zn(II)/m²] were loaded on the membranes by changing the initial concentration of Zn(II) ions and pH. Bovine serum albumin (BSA) adsorption on these membranes from aqueous solutions containing different amounts of BSA at different pH was investigated in batch reactors. The nonspecific adsorption of BSA on the poly(HEMA) membranes was negligible. Cibacron Blue F3GA attachment significantly increased the BSA adsorption up to 92.1 mg BSA/m². Adsorption capacity was further increased when Zn(II) ions were attached (up to 144.8 mg BSA m²). More than 90% of the adsorbed BSA was desorbed in 1 h in the desorption medium containing 0.5M NaSCN at pH 8.0 and 0.025M EDTA at pH 4.9. © 1998 John Wiley & Sons, Inc. J Appl Polym Sci 68: 657–664, 1998     

Human serum albumin adsorption on poly[(glycidyl methacrylate)‐co‐(methyl methacrylate)] beads modified with a spacer‐arm‐attached L‐histidine ligand

Poly(GMA/MMA) beads were synthesized from glycidyl methacrylate (GMA) and methyl methacrylate (MMA) in the presence of a cross‐linker (i.e. ethyleneglycol dimethacrylate) (EGDMA) via suspension polymerization. The epoxy groups of the poly(GMA/MMA) beads were converted into amino groups with either ammonia or 1,6‐diaminohexane (i.e. spacer‐arm). An L ‐histidine ligand was then covalently immobilized on the aminated (poly(GMA/MMA)‐AH) and/or the spacer‐arm attached (poly(GMA/MMA)‐SAH) beads using glutaric dialdehyde as a coupling agent. Both affinity adsorbents were used in human serum albumin (HSA) adsorption/desorption studies under defined pH, ionic strength or temperature conditions in a batch reactor. The spacer‐arm attached affinity adsorbent resulted in an increase in the adsorption capacity to HSA when compared to the aminated counterpart (i.e. poly(GMA/MMA)‐AH). The maximum adsorption capacities of the affinity adsorbents were found to be significantly high, i.e. 43.7 and 80.2 mg g^?1 (of the beads), while the affinity constants, evaluated by the Langmuir model, were 3.96 × 10^?7 and 9.53 × 10^?7 mol L^?1 for poly(GMA/MMA)‐AH and poly(GMA/MMA)‐SAH, respectively. The adsorption capacities of the affinity adsorbents were decreased for HSA by increasing the ionic strength, adjusted with NaCl. The adsorption kinetics of HSA were analysed by using pseudo‐first and pseudo‐second‐order equations. The second‐order equation fitted well with the experimental data. Copyright © 2005 Society of Chemical Industry     

Studies on accumulation of uranium by fungus Lentinus sajor-caju

The untreated, heat- and alkali-treated Lentinus sajor-caju mycelia were used for the recovery of uranium from aqueous solutions. The effect of pH, temperature, initial concentration of UO(2)(2+) ions and contact time parameters were investigated in a batch system. The particles sizes of the fungal mycelia were ranging from 100 to 200 microm. Biosorption equilibriums were established in about 30 min and the correlation regression coefficients show that the adsorption process can be well defined by the Freundlich equation. The alkali treated form had a high biosorption capacity (378 mg/g) than those of the untreated (268 mg/g) and heat-treated fungal mycelia (342 mg/g). Optimum biosorption was observed at pH 4.5 for all the tested fungal preparations and was independent of temperature (5-35 degrees C). In addition, the polarity and surface energy of the fungal biomass film preparations were determined by contact angle measurement. The fungal biomass could be regenerated using 10mM sodium carbonate, with up to 93% recovery. The biosorbents were used in six biosorption-desorption cycles and no considerable loss in the biosorption capacity was observed.     

Adsorption of Congo Red dye by native amine and carboxyl modified biomass of <Emphasis Type="Italic">Funalia trogii</Emphasis>: Isotherms,kinetics and thermodynamics mechanisms

Native, iminodiacetic acid and triethylenetetraamine modified biomasses of Funalia trogii were used for removal of Congo Red dye (CRD) from aqueous medium. The native and modified fungal biomasses were characterized using ATR-FTIR, Zeta potential, contact angle studies and analytical methods. FTIR studies of the native and chemically modified adsorbent preparations show that amine, carboxyl and hydroxyl groups are involved in the adsorption of the model dye (i.e., Congo Red). The maximum adsorption of the CRD on the native, carboxyl and amine groups modified fungal biomasses was obtained at pH 5.0. The amount of adsorbed dye on the adsorbent samples increased as the initial concentration of CRD in the solution increased to 200mg/L. The adsorption capacities of native, carboxyl groups and amine modified fungal preparations were 90.4, 153.6 and 193.7mg/g dry adsorbents, respectively. The data was fitted well with the Langmuir isotherm model, and followed the pseudo-second-order equations. Thermodynamic parameters (ΔG ^o , ΔH ^o and ΔS ^o ) were also calculated. The results showed that triethylenetetraamine (TETA) modified biomass of F. trogii presented an excellent dye removal performance and can be used in various environmental applications such as various micro-pollutants removal from aqueous medium.     

Kinetics of mercury ions removal from synthetic aqueous solutions using by novel magnetic p(GMA-MMA-EGDMA) beads

Poly(glycidylmethacrylate-methylmethacrylate), p(GMA-MMA-EGDMA), magnetic beads were prepared via suspension polymerization in the presence of ferric ions. The epoxy groups of the beads were converted into amino groups via ring opening reaction of the ammonia and, the aminated magnetic beads were used for the removal of Hg(II) ions from aqueous solution in a batch experiment and in a magnetically stabilized fluidized bed reactor (MFB). The magnetic p(GMA-MMA-EGDMA) beads were characterized with scanning electron microscope (SEM), FT-IR and ESR spectrophotometers. The optimum removal of Hg(II) ions was observed at pH 5.5. The maximum adsorption capacity of Hg(II) ions by using the magnetic beads was 124.8+/-2.1 mgg(-1) beads. In the continuous MFB reactor, Hg(II) ions adsorption capacity of the magnetic beads decreased with an increase in the flow-rate. The maximum adsorption capacity of the magnetic beads in the MFB reactor was 139.4+/-1.4 mgg(-1). The results indicate that the magnetic beads are promising for use in MFB for removal of Hg(II) ions from aqueous solution and/or waste water treatment.     

Epoxy‐derived pHEMA membrane for use bioactive macromolecules immobilization: Covalently bound urease in a continuous model system

Poly(2‐hydroxyethylmethacrylate) (pHEMA) membranes were prepared by UV‐initiated photopolymerization of HEMA in the presence of an initiator (α‐α′‐azobisisobutyronitrile, AIBN). The epoxy group, i.e., epichlorohydrin, was incorporated covalently, and the urease was immobilized onto pHEMA membranes by covalent bonding through the epoxy group. The retained activity of the immobilized enzyme was found to be 27%. The K_m values were 18 and 34 mM for the free and the immobilized enzymes, respectively, and the V_max values were found to be 59.7 and 16.2 U mg⁻¹ for the free and the immobilized enzyme. The optimum pHs was 7.2 for both forms, and the optimum temperature for the free and the immobilized enzymes were determined to be 45 and 50°C, respectively. The immobilized urease was characterized in a continuous system and during urea degradation the operational stability rate constant for the immobilized enzyme was found to be 5.83 × 10⁻⁵ min⁻¹. © 2000 John Wiley & Sons, Inc. J Appl Polym Sci 77: 2000–2008, 2000     

Adsorption of trypsin onto magnetic ion‐exchange beads of poly(glycidylmethacrylate‐co‐ethyleneglycoldimethacrylate)

Magnetic beads were prepared from glycidyl methacrylate (GMA), and ethyleneglycol dimethylmethacrylate (EGDMA) in the presence of Fe₃O₄ nano‐powder via suspension polymerization. The magnetic beads were characterized by surface area measurement, electron spin resonance (ESR), and scanning electron microscopy (SEM). ESR data revealed that the beads were highly super‐paramagnetic. The effects of contact time, pH, ionic strength, and temperature on the adsorption process have been studied. Adsorption equilibrium was established in about 120 min. The maximum adsorption of trypsin on the magnetic beads was obtained as 84.96 mg g^?1 at around pH 7.0. At increased ionic strength, the contribution of the electrostatic component to the overall binding decreased, and so the adsorption capacity. The experimental equilibrium data obtained trypsin adsorption onto magnetic beads fitted well to the Langmuir isotherm model. The result of kinetic analyzed for trypsin adsorption onto magnetic ion‐exchange beads showed that the second order rate equation was favorable. It was observed that after six adsorption–elution cycle, magnetic beads can be used without significant loss in trypsin adsorption capacity. Finally, the magnetic beads were used for separation of bovine serum albumin (BSA) and trypsin from binary solution in a batch system. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci, 2008