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PubMed2004

Characterisation and quality assessment of binding sites on a propazine-imprinted polymer prepared by precipitation polymerisation.

In this paper, the Langmuir-Freundlich isotherm (LF) is used to characterise a propazine-imprinted polymer obtained by precipitation polymerisation (MIP-P). Different rebinding studies were carried out allowing to explain the different interactions taking place between the molecularly imprinted polymer and six triazinic herbicides (desisopropylatrazine, desethylatrazine, simazine, atrazine, propazine and prometryn). The LF fitting parameters obtained (total number of binding sites, heterogeneity index and mean binding affinity) were compared to those obtained in a previous work for a propazine-imprinted polymer prepared by bulk polymerisation (MIP-B). From that study, it was concluded that precipitation polymerisation yielded polymers with a more homogeneous binding site distribution and higher affinity constants.

Binding Sites↗
PubMed2006

Evaluation of the influence of protein precipitation prior to on-line SPE-LC-API/MS procedures using multivariate data analysis.

Matrix effects on mass spectrometry (MS) response were investigated with three atmospheric pressure ionization (API) sources after on-line solid-phase extraction (SPE) of human plasma. On-line SPE was evaluated with one restricted access material (RAM), two large particle supports (LPS) and one monolith. A sample protein precipitation (PP) with acetonitrile (2:1) and a direct injection were tested. Principal component analysis (PCA) was performed to simplify data presentation and interpretation. Protein precipitation was found to be mandatory for reducing signal modification. Regarding sensitivity towards matrix effects after PP, atmospheric pressure photoionization (APPI) was globally the least sensitive ionization mode while electrospray ionization ESI was the most sensitive.

Alanine↗
PubMed2006

Sulfate adsorption and surface precipitation on a volcanic ash soil (allophanic andisol).

Sulfate strongly adsorbs on metal oxides and soils with variable charges. However, its surface precipitation has not been clearly evaluated and its adsorption mechanism has been in dispute. In the present study, an allophanic andisol, a typical volcanic ash soil having both negative and positive variable charges, was used to identify the adsorption mechanism of sulfate. Sulfate adsorption isotherms were obtained by a batch method at pH values of 4, 5, 6, and 7 in a wide range of concentrations in an Na-H-SO(4)-OH system. Theoretical isotherms were applied to the measured values for the evaluation. The surface precipitation was detected by the measured adsorption isotherms, and the BET isotherm confirmed the presence of multilayer adsorption. Stronger and weaker adsorption sites were suggested by using the Langmuir isotherm for the monolayer adsorption. The adsorption energies obtained from the Langmuir equation and recent spectroscopic analysis suggested that the stronger adsorption corresponded to an inner-sphere surface complex and that the weaker adsorption corresponded to outer-sphere surface complexation. The BET and Langmuir equations showed three types of adsorption mechanisms for the sulfate adsorption on the soil.

Adsorption↗
PubMed2005

Precipitation casting of drug-loaded microporous PCL matrices: incorporation of progesterone by co-dissolution.

Microporous, poly(epsilon-caprolactone) (PCL) matrices were loaded with progesterone by precipitation casting using co-solutions of PCL and progesterone in acetone. Progesterone loadings up to 32% w/w were readily achieved by increasing the drug content of the starting PCL solution. The kinetics of steroid release in PBS at 37 degrees C over 10 days could be described effectively by a diffusional release model although the Korsmeyer-Peppas model indicated the involvement of multiple release phenomena. The diffusion rate constant (D) increased from 8 to 24 microg/mg matrix/day0.5 as the drug loading increased from 3.6 to 12.4% w/w. A total cumulative release of 75%-95% indicates the high efficiency of steroid delivery. Increasing the matrix density from 0.22 to 0.39 g/cm3, by increasing the starting PCL solution concentration, was less effective in changing drug release kinetics. Retention of anti-proliferative activity of released steroid was confirmed using cultures of breast cancer epithelial (MCF-7) cells. Progesterone released from PCL matrices into PBS at 37 degrees C over 14 days retarded the growth of MCF-7 cells by a factor of at least 3.5 compared with progesterone-free controls. These findings recommend further investigation of precipitation-cast PCL matrices for delivery of bioactive molecules such as anti-proliferative agents from implanted, inserted or topical devices.

Animals↗
PubMed2004

Recovery of EDTA from complex solution using Cu(II) as precipitant and Cu(II) subsequent removal by electrolysis.

Ethylendiaminetetraacetate (EDTA) is a chelating agent widely used in industry and agriculture. Resistant to chemical and biological degradation EDTA represents a serious ecological problem. In order to avoid the outlet into the environment a new method of EDTA recycling has been proposed. The method involves substituting of the metal ions in EDTA complexes by Cu(II) and formation of an insoluble Cu2EDTA.4H2O compound at the excess of Cu(II) ions in weakly acidic solutions. Cu(II) ions substitute such metal ions as Ni(II), Zn(II), Co(II), Cd(II), Ca(II) and Mg(II). After treatment of the precipitate with water only, acidic or alkaline solutions the copper from the suspension formed can be removed by electrolysis. The highest current efficiency under galvanostatic conditions is in alkaline solutions, however, the highest yield of EDTA recovery is in acidic solutions. FT-IR investigations and chemical analysis of the precipitate formed have shown that in acidic and in alkaline solutions, H4EDTA and Na2H2EDTA.2H2O were formed, respectively. Electrolysis in acidic solutions gives the best results, i.e. the formed H4EDTA contains the highest amount of EDTA (95%) and the lowest amount of copper (0.01%).

Chelating Agents↗
PubMed2005

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.

Chemical Precipitation↗
PubMed2005

Treatment of organophosphate-contaminated wastewater by acidic hydrolysis and precipitation.

Wastewater from commercial manufacture of organophosphate (OP) pesticide of O-methyl-O-(2-isopropyl salicylate) thiophosphorusyl amide contains large amounts of organophosphate, organic sulfides, COD and NH3-N. A treatment process including acidic hydrolysis and precipitation was explored in this paper. The effects of initial pH values, hydrolysis temperature and hydrolysis time on the removal of pollutants were investigated. The experimental results showed that, in the hydrolysis, with the decreasing initial pH value, the removal of OP, sulfide and COD rose whereas the NH3-N removal declined. Increased hydrolysis temperature and time favored the removal of all the pollutants. 15.0 wt% Ca(OH)2 was sufficient for the removal of inorganic phosphorus and residual NH3-N in the precipitation step. With this chemical process, the removal of total phosphorus (TP), organophosphate, sulfides and NH3-N exceeded 90%, and the removal of COD was about 60%. The BOD5/COD value was greatly improved from 0.05 to 0.3, and the bio-degradability of the wastewater was greatly improved. The NH3 generated in the process was absorbed with water and might be reused in the manufacture of the pesticide. The results showed that this comprehensive process is effective for the treatment of this typical unbio-degradable pesticide wastewater.

Chemical Precipitation↗
PubMed2005

Enhanced sensitivity of surface plasmon resonance (SPR) immunoassays using a peroxidase-catalyzed precipitation reaction and its application to a protein microarray.

We describe a method to improve the sensitivity of surface plasmon resonance (SPR) immunoassays using a horseradish peroxidase (HRP)-catalyzed precipitation reaction. The precipitation reaction catalyzed by HRP bound to the SPR biosensor surface via a sandwich immunoassay induced a shift in the SPR angle. Human interferon (IFN)-gamma at concentrations ranging from 0.01 to 100 ng/ml was detectable by this method. We also show that this biocatalytic signal amplification method can be applied to SPR imaging (SPRI), in an immunoassay of multiple proteins on a protein microarray format.

Catalysis↗
PubMed2004

Protein precipitation for the analysis of a drug cocktail in plasma by LC-ESI-MS.

Three protein precipitation (PP) procedures with acetonitrile (ACN), perchloric acid (PA) and trichloroacetic acid (TCA) were investigated for the analysis of a drug cocktail from human plasma samples containing three pharmaceutical compounds and their primary metabolites. For this purpose, a capillary liquid chromatography-electrospray ionisation-mass spectrometry (LC-ESI-MS) method was developed for the simultaneous analysis of the six tested compounds in less than 6 min. Matrix effect was tested for each PP procedure by means of a post-column infusion system. The three PP techniques were found effective in removing proteins from human plasma and were fully compatible with capillary LC-ESI-MS analysis. However, with acid precipitations, low analyte recovery and a high variability, probably due to analyte coprecipitation, were obtained. Finally, ACN was found to be the most effective PP technique with a recovery higher than 80% and CV inferior to 6%.

Blood Proteins↗
PubMed2005

Precipitation of enzyme-catalyzed phenol oxidative coupling products: background ion and pH effects.

The effects of solution pH and background ion types and concentrations on the precipitation of polymeric products generated in the catalytically facilitated oxidative coupling of phenol were investigated systematically. The coupling reactions mediated by horseradish peroxidase were carried out under a specific predetermined experimental condition. Acids, bases, and/or selected salts of ions having different valences were then added to the resulting product solutions to adjust pH and ionic conditions. Subsequent analyses of product distributions between dissolved and precipitated forms revealed that ionic conditions and pH significantly impact those distributions, and associated mechanisms are discussed. The findings reported will assist feasibility assessments and process optimization with respect to engineering applications of catalyzed oxidative coupling reactions for wastewater treatment and soil decontamination.

Catalysis↗
PubMed1998

Precipitation of hard exudate after resorption of intraretinal edema after treatment of retinal branch vein occlusion.

PURPOSE: To present evidence that precipitation of hard exudate in the retina can occur after reduction of macular edema by successful photocoagulation treatment for retinal branch vein occlusion. METHODS: A 59-year-old woman received argon laser retinal photocoagulation treatment for retinal branch vein occlusion over the primarily involved engorged area of the retina. Fundus changes were documented by fundus photography and optical coherence tomography. RESULTS: Before treatment, an extensive intraretinal edema with cavity formation and a small serous foveal detachment extended outside the engorged area of the retina. Three months after treatment, all retinal thickening had disappeared, but a markedly increased amount of intraretinal hard exudate was seen adjacent to the treated area. CONCLUSIONS: Precipitation of hard exudate can occur after reduction of macular edema by successful retinal photocoagulation treatment, presumably because water and highly soluble electrolytes are resorbed faster than lipids. This phenomenon should be distinguished from accumulation of hard exudate resulting from disease progression.

Chemical Precipitation↗
PubMed1998

Functional reconstitution of membrane glycoproteins into lipid vesicles using lectin precipitation. Application to the VIP receptor.

We studied the interaction of n-octyl-beta-d-glucopyranoside-solubilized VIP receptors (VIPR) with wheat germ agglutinin and found that the addition of the lectin to the detergent extract led to the formation of aggregates that could be pelleted by high speed centrifugation. Resuspension of the pellet in the presence of the competing trisaccharide, N,N', N"-triacetylchitotriose (TAC), dissociated the lectin from the complex without altering the precipitability of VIPR. The final pellet (referred to as TAC pellet) contained an average of 12% of total protein and 96% of total VIP binding activity with a typical rank order of potency for VIP-related peptides. Lipid analysis and electron microscopic examination indicated that the precipitated material was composed of lipid vesicles. VIPR molecules were shown to be integrally inserted in the liposomes because they could not be dissociated from the vesicles at pH 11 or with high salt concentration. By comparing the liposome-associated VIP binding activity in the presence and absence of detergent and by showing accessibility of VIPR to PNGase F, it was concluded that VIP binding sites were not simply trapped within the reconstituted vesicles but likely exposed at the external surface of the liposomes.

Amidohydrolases↗
PubMed2003

Differential influence of proteolysis by calpain 2 and Lp82 on in vitro precipitation of mouse lens crystallins.

The purpose of the present study was to compare the susceptibility of crystallins proteolyzed by ubiquitous calpain 2 and by lens-specific calpain Lp82 to insolubilization. To test this, transgenic (TG) mice expressing a calpain 2, in which the active site cysteine 105 was mutated to alanine, were produced. Expression of mutated calpain 2 was driven in lens by coupling the mutated gene to the betaB1-crystallin promoter. Light scattering was measured in solutions of lens proteins after activation of endogenous calpain 2 and/or Lp82. Mass spectrometric analysis was performed to determine the cleavage sites and the calpain responsible for insolubilization of crystallins. Lens proteins from TG mice incubated in vitro with calcium showed higher light scattering compared to proteins from wild type (WT) mice. alphaA-crystallin from TG mice was proteolyzed by Lp82. In contrast, alphaA-crystallin in lenses from WT mice were proteolyzed by both calpain 2 and Lp82. These results suggested that Lp82-induced proteolysis of crystallins caused increased susceptibility of truncated crystallins to in vitro precipitation. Since Lp82 is highest in young animals, Lp82-induced proteolysis and precipitation may be one of the factors responsible for the cataract formation in young rodents.

Animals↗
PubMed2000

Chemical precipitation of apolipoprotein B-containing lipoproteins facilitates determination of LDL particle size.

OBJECTIVES: To simplify the determination of low-density lipoprotein (LDL) particle size by eliminating the need for ultracentrifugation. DESIGN AND METHODS: We compared LDL particle size determination by gradient gel electrophoresis using two different methods for separation of LDL: (a) by ultracentrifugation with a density between 1.019 and 1.063 g/mL, and (b) by precipitation of the apolipoprotein B-containing lipoproteins from plasma. LDL particle size was determined for 41 individuals using both methods. RESULTS: The correlation between these two methods was r = 0.98; peak particle diameter (nm) was reproducible with a coefficient of variation of 1. 3% for LDL separated by ultracentrifugation and 1.4% for LDL prepared by precipitation. The intra-assay variation within a single gel was 0.2%. CONCLUSION: Elimination of the need for ultracentrifugation or lipid staining significantly reduces the cost and simplifies the procedure of LDL particle sizing. As a result, larger patient populations can be more readily screened for the determination of LDL particle size.

Adult↗
PubMed2002

Salt-precipitation method does not isolate to homogeneity Tamm-Horsfall glycoprotein from urine of proteinuric patients and pregnant women.

OBJECTIVE: Assessment of the degree of purification of Tamm-Horsfall glycoprotein from anomalous urine. DESIGN AND METHODS: Two methods have been compared: the method of Tamm & Horsfall (T&H method) consisting in the precipitation of THP by the addition to urine of NaCl up to 0.58 mol/L and the filtration of urine through a diatomaceous earth filter (DEF method) in which THP is selectively trapped because of its gelation/aggregation tendency. The purity of THP preparations has been evaluated by SDS-PAGE analysis and Western blotting developed with anti immunoglobulin G (IgG) antibodies and antichorionic gonadotropin antibodies. RESULTS: All THPs isolated by T&H method from proteinuric patients were contaminated by IgG and one of the five preparations from pregnant women even by chorionic gonadotropin. A smaller or no contamination was found in THPs isolated by DEF method. CONCLUSIONS: Although albumin is the most abundant protein in the anomalous urine, it never appears in THP preparations. The consistent contamination with IgG of THP prepared by salt precipitation-method might be related to the formation of a stable complex between the two proteins.

Animals↗
PubMed1991

Precipitation method for separating and quantifying bone and liver alkaline phosphatase isoenzymes.

We evaluated a method for quantifying bone isoenzyme of alkaline phosphatase (ALP) which utilizes wheat-germ lectin to precipitate this fraction. In precision studies, CVs ranged from 3.2 to 11.4% (within-day) and from 3.7 to 11.5% (day-to-day). The assay procedure was linear to 1100 U/L and was easily adapted to automated kinetic measurement. Comparison of the precipitation method with an affinity electrophoretic method, which utilizes cellulose acetate as a support, demonstrated a satisfactory coefficient of correlation (r = 0.886). The reference range was determined in sera from 188 healthy adult subjects. The distribution of bone ALP values was also studied in 73 healthy children and in 30 healthy adolescents. To evaluate the clinical applicability of the method, the bone isoenzyme was determined in samples from several groups of subjects (pregnant women, patients with hepatobiliary diseases, patients with hepatocellular carcinoma without skeletal involvement, and patients with bone, liver or lymph node metastases). We found the method suitable for routine determination of bone alkaline phosphatase and for the screening of bone metastases. Because of its technical simplicity and satisfactory analytical performance, it can be used instead of the heat-inactivation procedure.

Adolescent↗
PubMed1985

Precipitation methods for the determination of LDL-cholesterol.

The selective precipitation of low-density lipoproteins (LDL) with polyvinyl sulfate (PVS), and the immunoprecipitation of high-density lipoproteins (HDL) and very-low-density lipoproteins (VLDL) with an anti-HDL antibody, can both be used to establish simple methods for the determination of LDL cholesterol. Whereas the PVS method requires the calculation of LDL cholesterol as the difference of total and supernatant cholesterol, the immunoprecipitation method allows the direct measurement of LDL cholesterol in the supernatant. As a first step, both methods were optimized to yield accurate values for normolipemic and slightly hyperlipemic serum samples. Moreover, the determination of LDL-cholesterol in lipemic sera can be achieved by a combination of immunoprecipitation and polyanion precipitation.

Chemical Precipitation↗
PubMed1977

Serological precipitation method for studying biosynthesis and secretion of immunoglobulins by human peripheral blood lymphocytes.

A reproducible, serological method to quantify radiolabeled immunoglobulin (Ig) synthesized and secreted by human peripheral blood lymphocytes is described. In contrast to indirect precipitation (rabbit antihuman Ig and goat antirabbit Ig), the direct coprecipitation method using human IgG and rabbit antihuman IgG F (ab)2 showed much smaller, non-specifically precipitable radioactivity (i.e., 2 approximately 14%) without affecting Ig radioactivity.

Animals↗