Optimization of alginate purification using polyvinylidene difluoride membrane filtration: Effects on immunogenicity and biocompatibility of three-dimensional alginate scaffolds.

Sodium alginate is an effective biomaterial for tissue engineering applications. Non-purified alginate is contaminated with protein, lipopolysaccharide, DNA, and RNA, which could elicit adverse immunological reactions. We developed a purification protocol to generate biocompatible alginate based on (a) activated charcoal treatment, (b) use of hydrophobic membrane filtration (we used hydrophobic polyvinylidene difluoride membranes to remove organic contaminants), (c) dialysis, and finally (d) ethanol precipitation. Using this approach, we could omit pre-treatment with chloroform and significantly reduce the quantities of reagents used. Purification resulted in reduction of residual protein by 70% down to 0.315 mg/g, DNA by 62% down to 1.28 µg/g, and RNA by 61% down to less than 10 µg/g, respectively. Lipopolysaccharide levels were reduced by >90% to less than 125 EU/g. Purified alginate did not induce splenocyte proliferation in vitro. Three-dimensional scaffolds generated from purified alginate did not elicit a significant foreign body reaction, fibrotic overgrowth, or macrophage infiltration 4 weeks after implantation. This study describes a simplified and economical alginate purification method that results in alginate purity, which meets clinically useful criteria.

Extraction of alumina and sodium oxide from red mud by a mild hydro-chemical process.

A mild hydro-chemical process to extract Al(2)O(3) in red mud to produce sodium aluminate hydrate was investigated, and the optimum conditions of Al(2)O(3) extraction were verified by experiments as leaching in 45% NaOH solution with CaO-to-red mud mass ratio of 0.25 and liquid-to-solid ratio of 0.9, under 0.8 MPa at 200 degrees C for 3.5h. Subsequent process of extracting Na(2)O from the residue of Al(2)O(3) extraction was carried out in 7% NaOH solution with liquid-to-solid ratio of 3.8 under 0.9 MPa at 170 degrees C for 2h. Overall, 87.8% of Al(2)O(3) and 96.4% of Na(2)O were extracted from red mud. The final residues with less than 1% Na(2)O could be utilized as feedstock in construction materials. The chemical reactions taking place in both Al(2)O(3) and Na(2)O extractions from red mud are proposed.

Perchlorate reduction by autotrophic bacteria in the presence of zero-valent iron.

A series of batch experiments were performed to study the combination of zero-valent iron (ZVI) with perchlorate-reducing microorganisms (PRMs) to remove perchlorate from groundwater. In this method, H2 produced during the process of iron corrosion by water is used by PRMs as an electron donor to reduce perchlorate to chloride. Perchlorate degradation rates followed Monod kinetics, with a normalized maximum utilization rate (rmax) of 9200 microg g(-1) (dry wt) h(-1) and a half-velocity constant (Ks) of 8900 microg L(-1). The overall rate of perchlorate reduction was affected by the biomass density within the system. An increase in the OD600 from 0.025 to 0.08 led to a corresponding 4-fold increase of perchlorate reduction rate. PRM adaptation to the local environment and initiation of perchlorate reduction was rapid under neutral pH conditions. At the initial OD600 of 0.015, perchlorate reduction followed pseudo-first-order reaction rates with constants of 0.059 and 0.033 h(-1) at initial pH 7 and 8, respectively. Once perchlorate reduction was established, the bioreductive process was insensitive to the increases of pH from near neutral to 9.0. In the presence of nitrate, perchlorate reduction rate was reduced, but not inhibited completely.

A hybrid liquid-phase precipitation (LPP) process in conjunction with membrane distillation (MD) for the treatment of the INEEL sodium-bearing liquid waste.

A novel hybrid system combining liquid-phase precipitation (LPP) and membrane distillation (MD) is integrated for the treatment of the INEEL sodium-bearing liquid waste. The integrated system provides a "full separation" approach that consists of three main processing stages. The first stage is focused on the separation and recovery of nitric acid from the bulk of the waste stream using vacuum membrane distillation (VMD). In the second stage, polyvalent cations (mainly TRU elements and their fission products except cesium along with aluminum and other toxic metals) are separated from the bulk of monovalent anions and cations (dominantly sodium nitrate) by a front-end LPP. In the third stage, MD is used first to concentrate sodium nitrate to near saturation followed by a rear-end LPP to precipitate and separate sodium nitrate along with the remaining minor species from the bulk of the aqueous phase. The LPP-MD hybrid system uses a small amount of an additive and energy to carry out the treatment, addresses multiple critical species, extracts an economic value from some of waste species, generates minimal waste with suitable disposal paths, and offers rapid deployment. As such, the LPP-MD could be a valuable tool for multiple needs across the DOE complex where no effective or economic alternatives are available.

Hydrogen-based, hollow-fiber membrane biofilm reactor for reduction of perchlorate and other oxidized contaminants.

Many oxidized pollutants, such as nitrate, perchlorate, bromate, and chlorinated solvents, can be microbially reduced to less toxic or less soluble forms. For drinking water treatment, an electron donor must be added. Hydrogen is an ideal electron donor, as it is non-toxic, inexpensive, and sparsely soluble. We tested a hydrogen-based, hollow-fiber membrane biofilm reactor (MBfR) for reduction of perchlorate, bromate, chlorate, chlorite, chromate, selenate, selenite, and dichloromethane. The influent included 5 mg/L nitrate or 8 mg/L oxygen as a primary electron accepting substrate, plus 1 mg/L of the contaminant. The mixed-culture reactor was operated at a pH of 7 and with a 25 minute hydraulic detention time. High recirculation rates provided completely mixed conditions. The objective was to screen for the reduction of each contaminant. The tests were short-term, without allowing time for the reactor to adapt to the contaminants. Nitrate and oxygen were reduced by over 99 percent for all tests. Removals for the contaminants ranged from a minimum of 29% for chlorate to over 95% for bromate. Results show that the tested contaminants can be removed as secondary substrates in an MBfR, and that the MBfR may be suitable for treating these and other oxidized contaminants in drinking water.

Perchlorate removal in sand and plastic media bioreactors.

The treatment of perchlorate-contaminated groundwater was examined using two side-by-side pilot-scale fixed-bed bioreactors packed with sand or plastic media, and bioaugmented with the perchlorate-degrading bacterium Dechlorosoma sp. KJ. Groundwater containing perchlorate (77microg/L), nitrate (4mg-NO(3)/L), and dissolved oxygen (7.5mg/L) was amended with a carbon source (acetic acid) and nutrients (ammonium phosphate). Perchlorate was completely removed (<4microg/L) in the sand medium bioreactor at flow rates of 0.063-0.126L/s (1-2gpm or hydraulic loading rate of 0.34-0.68L/m(2)s) and in the plastic medium reactor at flow rates of <0.063L/s. Acetate in the sand reactor was removed from 43+/-8 to 13+/-8mg/L (after day 100), and nitrate was completely removed in the reactor (except day 159). A regular (weekly) backwashing cycle was necessary to achieve consistent reactor performance and avoid short-circuiting in the reactors. For example, the sand reactor detention time was 18min (hydraulic loading rate of 0.68L/m(2)s) immediately after backwashing, but it decreased to only 10min 1 week later. In the plastic medium bioreactor, the relative changes in detention time due to backwashing were smaller, typically changing from 60min before backwashing to 70min after backwashing. We found that detention times necessary for complete perchlorate removal were more typical of those expected for mixed cultures (10-18min) than those for the pure culture (<1min) reported in our previous laboratory studies. Analysis of intra-column perchlorate profiles revealed that there was simultaneous removal of dissolved oxygen, nitrate, and perchlorate, and that oxygen and nitrate removal was always complete prior to complete perchlorate removal. This study demonstrated for the first time in a pilot-scale system, that with regular backwashing cycles, fixed-bed bioreactors could be used to remove perchlorate in groundwater to a suitable level for drinking water.

Rate and extent of aqueous perchlorate removal by iron surfaces.

The rate and extent of perchlorate reduction on several types of iron metal was studied in batch and column reactors. Mass balances performed on the batch experiments indicate that perchlorate is initially sorbed to the iron surface, followed by a reduction to chloride. Perchlorate removal was proportional to the iron dosage in the batch reactors, with up to 66% removal in 336 h in the highest dosage system (1.25 g mL(-1)). Surface-normalized reaction rates among three commercial sources of iron filings were similar for acid-washed samples. The most significant perchlorate removal occurred in solutions with slightly acidic or near-neutral initial pH values. Surface mediation of the reaction is supported by the absence of reduction in batch experiments with soluble Fe2+ and also by the similarity in specific reaction rate constants (kSA) determined for three different iron types. Elevated soluble chloride concentrations significantly inhibited perchlorate reduction, and lower removal rates were observed for iron samples with higher amounts of background chloride contamination. Perchlorate reduction was not observed on electrolytic sources of iron or on a mixed-phase oxide (Fe3O4), suggesting that the reactive iron phase is neither pure zerovalent iron nor the mixed oxide alone. A mixed valence iron hydr(oxide) coating or a sorbed Fe2+ surface complex represent the most likely sites for the reaction. The observed reaction rates are too slow for immediate use in remediation system design, but the findings may provide a basis for future development of cost-effective abiotic perchlorate removal techniques.

Use of surfactant modified ultrafiltration for perchlorate (Cl(O)(4-)) removal.

Determinations of perchlorate anion (ClO(4)(-)) transport and rejection were performed using a surfactant modified ultrafiltration (UF) membrane. Perchlorate anion (at a concentration of 100 microg/L of ClO(4)(-), spiked with KClO(4)) was introduced to the membrane as a pure component, in binary mixtures with other salts, cationic and anionic surfactants, and at various ionic strength conditions (conductivity). Also, a natural source water was spiked with perchlorate in the presence of cationic and anionic surfactants and used to determine the effects of a complex mixture (including natural organic matter (NOM)) on the observed rejection. All filtration measurements were performed at approximately the same permeate flow rate in order to minimize artifacts from mass transfer at the membrane interface. The objective of this study was to modify a negatively charged UF membrane in terms of the fundamental mechanisms, steric/size exclusion and electrostatic exclusion and to enhance perchlorate rejection, with synthetic water and a blend of Colorado River water and State Project water (CRW/SPW). Previous work suggested that perchlorate was dominantly rejected by electrostatic exclusion for charged nanofiltration (NF) and UF membranes (Rejection of perchlorate by reverse osmosis, nanofiltration and ultrafiltration (UF) membranes: mechanism and modeling. Ph.D. dissertation, University of Colorado, Boulder, USA, 2001). In that research, perchlorate rejection capability was quickly lost in the presence of a sufficient amount of other ions. However, this study showed that ClO(4)(-) was excluded from a (negatively) charged UF membrane with pores large with respect to the size of the ion. Although perchlorate rejection capability due to apparent electrostatic force was reduced in the presence of a cationic surfactant, a desired amount of the ClO(4)(-) was excluded by steric exclusion. The steric exclusion was due to decreasing membrane pore size caused by the adsorption of the cationic surfactant.

The sensitivity of fixed-bed biological perchlorate removal to changes in operating conditions and water quality characteristics.

Flow rate, electron donor addition, and biomass control were evaluated in order to optimize perchlorate (ClO4-) removal from drinking water using biologically active carbon (BAC) filtration. Influent dissolved oxygen (DO) was lowered from ambient conditions to approximately 2.5 mg/L for all experiments using a nitrogen sparge. When influent nitrate concentration was 0-2.0 mg/L, 1.6-2.8 mg/L as carbon of acetate or ethanol was required to achieve and sustain the complete removal of 50 microg/L perchlorate in a BAC filter. Most or all of the exogenous acetate and ethanol was removed during biofiltration. When a 72-h electron donor feed failure was simulated, a maximum perchlorate breakthrough of 18 microg/L was observed and, once electron donor was reapplied, 9 days were required to reestablish complete perchlorate removal. During a 24-h electron donor feed failure simulation, the maximum effluent perchlorate concentration detected was 6.7 microg/L. Within 24 h of reactivating the electron donor, the filter regained its capacity to consistently remove 50 microg/L perchlorate to below detection. Although biomass growth diminished the filter's ability to consistently remove perchlorate, a cleaning procedure immediately restored stable, complete perchlorate removal. This cleaning procedure was required approximately every 50 days (4800 bed volumes) when influent DO concentration was 2.5 mg/L. Empty-bed contact time (EBCT) experiments showed that 80% perchlorate removal was achieved using a 5-min EBCT, and complete perchlorate removal was observed for an EBCT of 9 min. It was also demonstrated that BAC filtration consistently removed perchlorate to below detection for influent perchlorate concentrations ranging from 10 to 300 microg/L, influent sulfate concentrations between 0 and 220 mg/L, influent pH values of 6.5-9.0, and operating temperatures of 5-22 degrees C.

Treatment of perchlorate- and nitrate-contaminated groundwater in an autotrophic, gas phase, packed-bed bioreactor.

The biological degradation of perchlorate was examined using a laboratory-scale, autotrophic, packed-bed biofilm reactor. The reactor was operated in unsaturated-flow mode and continuously fed water containing perchlorate (ClO4-) (as an electron acceptor), and a gas mixture of hydrogen (5%) and carbon dioxide at a retention time of 1.5 min. In the absence of nitrate, perchlorate removal rate (rp, ppb/min) in the reactor was found to be first order with respect to perchlorate concentration (c, ppb) according to rp = 0.16 +/- 0.06c(0.97+/-0.12) (n = 11, R2 = 0.97, p < 10(-5)). Perchlorate removal rates in the hydrogen feed were found to be comparable to rates found by others for fixed film bioreactors using either hydrogen gas or organic electron donors such as acetate, although the rate coefficient was reduced to slightly less than unity (r(p) = 0.22 +/- 0.08c(0.91+/-0.08); n = 19, R2 = 0.89, p < 10(-5)). When nitrate was present in the water, similar perchlorate removals were achieved despite nitrate concentrations three orders of magnitude higher than perchlorate concentrations. Perchlorate was removed by an average of 25 +/- 5% from a perchlorate-contaminated groundwater containing 73 +/- 2 ppb of perchlorate and 21 +/- 2 ppm of nitrate. This removal was slightly higher than the removal of 17 +/- 3% measured for a synthetic groundwater containing 79 +/- 3 ppb of perchlorate and 22 +/- 2 ppm of nitrate. In both cases, there was an average of 10% nitrate removal.