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A mild HPLC method for the quality control of [18F]sodium fluoride solution for injection was developed. The method used anion exchange column for separation, UV and online radio-detectors for fluoride content and fluorine-18 determination. A mixture of 100 mM sodium acetate and 25 mM sodium chloride was chosen as mobile phase. The method was validated for system suitability, specificity, fluoride concentration and radiochemical purity determination with good linearity, accuracy and precision. The good robustness of the system was also verified, enabling routine analysis of fluoride content and radiochemical purity of the [18F]sodium fluoride product.

Phosphoinositide (PPI) lipids are a crucial class of low abundance signaling molecules that regulate many processes within cells. Methods that enable simultaneous detection of all PPI lipid species provide a wholistic snapshot of the PPI profile of cells, which is critical for probing PPI biology. Here we describe a method for the simultaneous measurement of cellular PPI levels by metabolically labeling yeast or mammalian cells with myo-(3)H-inositol, extracting radiolabeled glycerophosphoinositides, and separating lipid species on an anion exchange column via HPLC.

The stability of four major arsenic (As) species during application of the BARGE (Bioaccessibility Research Group of Europe) unified bioaccessibility method (UBM) has been assessed. The concentrations of As species in the UBM gastric and gastro-intestinal (gastric + intestinal) phases were determined using HPLC-ICP-MS whilst the total As content in the samples was determined using ICP-MS alone. The arsenic species studied were arsenite As(III), arsenate As(V), dimethylarsinic acid (DMA) and monomethylarsonic acid (MMA). These species were separated in 10 min using an anion exchange column (Hamilton PRP-X100) with a mobile phase containing 20 mmol L−1 NH4H2PO4/1% methanol (pH 6.0). The recoveries of arsenic species spiked into the gastric and gastro-intestinal fluids were in the range 90–108%. No interconversion between As species was observed as a result of applying the BARGE UBM, which is a particularly important finding for the reliability of As(III) measurements. The accuracy of the BARGE UBM for in vitro extractable As(V) was verified using British Geological Survey (BGS) guidance material 102 (an ironstone soil). For a commercial rice sample, the bioaccessibility sequence of As was DMA > As(III) > As(V) for the gastric phase and As(III) > DMA > As(V) for the gastro-intestinal phase.

In this study a new method for evaluating the pressure effect on separations of oligonucleotides and proteins on an anion exchange column was developed. The pressure rise of up to 500 bar was attained by coupling restriction capillaries to the column outlet to minimize differences in pressure over the column. Using pH transient measurements it was demonstrated that no shift in ion exchange equilibria occurs due to a pressure increase. Results from isocratic and gradient separations of oligonucleotides (model compounds) were evaluated by stoichiometric displacement and linear gradient elution model, respectively. Both elution modes demonstrated that for smaller oligonucleotides the number of binding sites remained unchanged with pressure rise while an increase for large oligonucleotides was observed, indicating their alignment over the stationary phase. From the obtained model parameters and their pressure dependencies, a thermodynamic description was made and compared between the elution modes. A complementary pattern of a linear increase of partial molar volume change with a pressure rise was established. Furthermore, estimation of the pressure effect was performed for bovine serum albumin and thyroglobulin that required gradient separations. Again, a raise in binding site number was found with pressure increase. The partial molar volume changes of BSA and Tg at the maximal investigated pressure and minimal salt concentration were −31.6 and −34.4 cm3/mol, respectively, indicating a higher rigidity of Tg. The proposed approach provides an insight into the molecule deformation over a surface at high pressures under nondenaturing conditions. The information enables a more comprehensive UHPLC method development.

Oligonucleotides (ONs) are widely used in different applications in life science, forensic, in i.e. family tree DNA test for humans and in diagnostic applications in several fields. The use of ONs in biopharmaceutical therapeutic areas also generates new challenges handling more complex molecules which results in the need of further developed analytical techniques. Anion exchange chromatography (AEX) is a common separation technique for biomolecules and is based on charge attraction between the analyte and the stationary phase. The chromatographic system is complex and often high pH and a high salt concentration is needed for the elution to occur, which in some systems can be corrosive for both the column and the instrument. The aim of this study was to evaluate new mobile phase compositions with lower salt concentrations, organic modifier, and usage of a buffer to increase the control of the pH. This was done by evaluation of three columns developed for AEX and uses different chemical and methodical modification of the mobile phase to control the retention to the stationary phase. The influence of pH, temperature, and methanol (MeOH) content in the buffer were studied by evaluation of resolution, asymmetry, and efficiency responses. Three oligonucleotides with 16, 18 and 19 T-bases in the chain were used in the study of three AIE columns. High pH, elevated temperature and the addition of an organic modifier were used for unfolding of the oligonucleotide chain and generating more efficient separations. Other parameters such as gradient slope and initial concentration of the eluting buffer were also studied, and the findings clearly show that the chromatographic conditions influence the resolution, asymmetry, and efficiency. Oligonukleotider används i flera olika branscher som forensik och inom life sience till diagnostiska och medicinska applikationer. Eftersom applikationsområdena ökar och forskningen går framåt så blir molekylerna mer komplexa, vilket i sin tur kräver att dom analytiska teknikerna också behöver utvecklas. En vanlig separationsteknik för biomolekyler är anjonbyteskromatografi, då den bygger på laddningsattraktion mellan analyten och den stationära fasen. Oligonukleotider har en negativ nettoladdning på grund av den negativt laddade fosfatgruppen i kedjan. Genom att modifiera de kemiska egenskaperna i den mobila fasen är det därför möjligt att i viss grad kontrollera retentionen till den stationära fasen. Då det kromatografiska systemet är komplext och det ofta behövs högt pH och/eller en hög saltkoncentration för eluering av analyten kan det i vissa system verka korrosivt för både kolonnen och instrumentet. Det initiala målet med detta arbete var att ta fram olika mobila faser med lägre saltkoncentration, organiskt lösningsmedel och användande av buffer för att kontrollera pH. Detta gjordes genom att utvärdera den kromatografiska kapaciteten hos tre olika anjonbyteskolonner samt använda olika kemiska- och metodiska modifieringar av den mobila fasen för att kontrollera analytens retention. Mer specifikt så kommer påverkan av pH, temperatur och innehåll av metanol i den mobila fasen att studeras genom att utvärdera upplösning, asymmetri och effektivitet i de erhållna kromatogrammen. Analytmixen innehåller tre olika oligonukleotider vilka består av 16, 18 eller 19 T-baser i sekvensen. Genom att höja pH, temperatur och innehåll av metanol i den mobila fasen påvisas mer effektiva separationer. Andra faktorer som gradientlutning och initialkoncentration av den eluerande fasen studeras också med goda resultat när det gäller dess påverkan på upplösning, asymmetri och effektivitet.

An evaluation was performed on the feasibility of using HB-Line anion exchange column waste streams from Alternate Feedstock 2 (AFS-2) processing for the dissolver solution for used nuclear fuel (UNF) processing. The targeted UNF for dissolution using recycled solution are fuels similar to the University of Missouri Research Reactor (MURR) fuel. The objectives of this experimental program were to validate the feasibility of using impure dissolver solutions with the MURR dissolution flowsheet to verify they would not significantly affect dissolution of the UNF in a detrimental manner.

Objective Management and diagnosis of multiple human cancers remains a challenge and search for a common biomarker is still debatable. In this manuscript we have evaluated the use of monoclonal antibody UNIVmAb, to detect the protein (H11) as a common biomarker for all cancers irrespective of the grade and origin. We have shown by both ELISA and Western Blot that the H11 protein, is a unique hyaluronan binding protein that has not been detected earlier. H11 protein was fractionated in an anion exchange column followed by cibacron blue gel exclusion chromatography. Hyaluronan binding H11 protein reacted with Monoclonal antibody UNIVmAb and b-HA inspite of b-Hyaluronan (biotinylated Hyaluronan) interaction and HA-Oligo (Hyaluronan oligosaccharides) competition from various grades of Human cancers sera. Results ELISA, Western blot and b-Hyaluronan interactions clearly showed an over-expression of UNIVmAb reacted H11 protein in all fifty cancer’s sera when compared with seventy normal sera. UNIVmAb reactive H11 protein can be used as a common biomarker. We believe, UNIVmAb detected H11 protein, is a unique hyaluronan binding protein, that can be used as a common biomarker for all cancers.

Emerging interest in extending the plasma half-life of small molecule radioligands warrants a consideration of the appropriate radionuclide for PET imaging at longer time points (&gt, 8 h). Among candidate positron-emitting radionuclides, 66Ga (t1/2 = 9.5 h, &beta, + = 57%) has suitable nuclear and chemical properties for the labeling and PET imaging of radioligands of this profile. We investigated the value of 66Ga to preclinical screening and the evaluation of albumin-binding PSMA-targeting small molecules. 66Ga was produced by irradiation of a natZn target. 66Ga3+ ions were separated from Zn2+ ions by an optimized UTEVA anion exchange column that retained 99.99987% of Zn2+ ions and allowed 90.2 &plusmn, 2.8% recovery of 66Ga3+. Three ligands were radiolabeled in 46.4 &plusmn, 20.5%, radiochemical yield and &gt, 90% radiochemical purity. Molar activity was 632 &plusmn, 380 MBq/&micro, mol. Uptake in the tumor and kidneys at 1, 3, 6, and 24 h p.i. was determined by &micro, PET/CT imaging and more completely predicted the distribution kinetics than uptake of the [68Ga]Ga-labeled ligands did. Although there are multiple challenges to the use of 66Ga for clinical PET imaging, it can be a valuable research tool for ligand screening and preclinical imaging beyond 24 h.

An analytical method that combined high-performance liquid chromatography with inductively coupled plasma mass spectrometry has been developed for the determination of hexavalent chromium in traditional Chinese medicines. Hexavalent chromium was extracted using the alkaline solution. The parameters such as the concentration of alkaline and the extraction temperature have been optimized to minimize the interconversion between trivalent chromium and hexavalent chromium. The extracted hexavalent chromium was separated on a weak anion exchange column in isocratic mode, followed by inductively coupled plasma mass spectrometry determination. To obtain a better chromatographic resolution and sensitivity, 75 mM NH4 NO3 at pH 7 was selected as the mobile phase. The linearity of the proposed method was investigated in the range of 0.2-5.0 μg L(-1) (r(2) = 0.9999) for hexavalent chromium. The limits of detection and quantitation are 0.1 and 0.3 μg L(-1) , respectively. The developed method was successfully applied to the determination of hexavalent chromium in Chloriti lapis and Lumbricus with satisfactory recoveries of 95.8-112.8%.

Methods for producing prothrombin complex concentrate (PPSB) by anion-exchange chromatography in batch mode using various sorbents such as DEAE- and QAE-Sephadex A-50 were examined. It was established that the yield and specific activity of FII, FVII, FIX, and FX depended on the mass of resin used for chromatography. New column chromatography methods for producing PPSB with a high FVII content were proposed. The static capacity of several sorbents for coagulation system FVII and the dynamic capacity of TOYOPEARL GigaCap Q-650M and Capto Q were studied.

A sensitive and accurate method based on ion chromatography was established for determination of five lithium salts in lithium-ion batteries electrolytes. Chromatographic analyses were carried out on an anion exchange column at flow rate of 1 mL/min. Under the optimal conditions, five target anions (BF 4 − , PF 6 − , TFSI − , BOB − and FSI − ) exhibited satisfactory linearity with a correlation coefficient of 0.9996. The relative standard derivations of the target anions were less than less than 0.94% ( n = 7). The limits of detections were in the range of 0.068–0.29 mg/L with average spiked recoveries ranging from 96.8% to 105.1%.

A comprehensive method for the precise determination of Re, Os, Ir, Ru, Pt and Pd concentrations as well as Os isotopic compositions in geological samples is presented. Samples were digested by the Carius tube method, and the Os was extracted by conventional CCl4 method. The Re, Ir, Ru, Pt and Pd were first subgroup separated from the matrix elements into Re-Ru, Ir-Pt and Pd by a 2-ml anion exchange column. Subsequently, the Re-Ru was further purified by a secondary 0.25 ml anion exchange column or by microdistillation of Ru using CrO3-H2SO4 as an oxidant followed by a secondary 0.25 ml anion exchange separation of Re. The Pd and Ir-Pt were further successively purified by an Eichrom-LN column to completely remove Zr and Hf, respectively. Rhenium, Ir, Ru, Pt and Pd were individually measured by multi-collector inductively coupled plasma-mass spectrometry (MC-ICP-MS), except for Ru after microdistillation purification was analysed by negative-thermal ionisation mass spectrometry (N-TIMS). The analytical results for peridotite reference material WPR-1 agree well with the previously published data. Finally, several mafic rock reference materials including TDB-1, WGB-1, BHVO-2, BCR-2, BIR-1a and DNC-1a were analysed for Re-Os isotopes and platinum-group element concentrations to test their suitability for certification. Une methode complete pour la determination precise des concentrations en Re, Os, Ir, Ru, Pt et Pd, ainsi que des compositions isotopiques l'Os dans des echantillons geologiques est presentee. Les echantillons ont ete digeres par la methode du tube de Carius et l'Os a ete extrait par la methode classique CCl4. Le Re, l'Ir, le Ru, le Pt et le Pd sont d'abord separes des autres elements de la matrice sous la forme Re-Ru, Ir-Pt et Pd grâce a une colonne d’echange d'anions de 2 ml. Ensuite, l'association Re-Ru est purifiee une seconde fois grâce a une colonne secondaire echangeuse d'anions de 0.25 ml, ou par une micro-distillation du Ru en utilisant l'association CrO3-H2SO4 comme oxydant, suivie par une separation secondaire du Re sur une colonne echangeuse d'anions de 0.25 ml. Le Pd et le couple Ir-Pt ont ete purifies successivement une seconde fois grâce a une colonne Eichrom-LN pour enlever totalement et successivement le Zr et l'Hf. Le rhenium, l'Ir, le Ru, le Pt et le Pd sont mesures individuellement par spectrometrie de masse a source plasma et a multi-collection (MC-ICP-MS), a l'exception du Ru qui apres purification par micro-distillation a ete analyse par spectrometrie de masse a thermo-ionisation negative (N-TIMS). Les resultats d'analyse de la peridotite de reference WPR-1 sont en bon accord avec les donnees publiees anterieurement. Enfin, plusieurs roches mafiques de reference dont TDB-1, WG B-1, BHVO-2, BCR-2, BIR-1a DNC-1a ont ete analysees pour les isotopes Re-Os et les concentrations en elements du groupe du platine pour tester la possibilite qu'elles puissent etre certifie.

A bacterial strain was isolated from Pasinler hot spring, Erzurum, Turkey. The purified thermophilic isolate was identified as Geobacillus pallidus P26 and used to produce extracellular pectin lyase (EC 4.2.2.10). Pectin lyase enzyme was purified 34 fold by using DEAE-cellulose anion exchange column chromatography and characterized. Molecular weight of the enzyme was determined as 56 kDa by using Sephadex G-100 gel filtration chromatography. Purification of enzyme was verified by SDS-PAGE. The pH- and temperature optima of enzyme were determined (pH 9.0 and 60°C, respectively). Pectin lyase was mostly stable at 50 oC for 24 hours. Its’ activity decreased to 50 % for 24 h at 60°C. KM and Vmax were calculated as 24.8 mg/mL and 2.28 μmol/L min, respectively. Purified pectin lyase was inhibited by Fe3+, Zn2+, Cu2+, Ca2+, Co2+ and Hg2+ but not by Mg2+. The purified pectin lyase enzyme was used for getting fruits juices. It was found that yields of fruits juices increased when they were compared with control.

The radionuclide 55Co (T1/2 = 17.5 h, Eβ+ = 1.5 MeV, 76% β+ decay) is an important β+ emitting radioisotope. For production of 55Co via natFe(p, xn) 55Co reaction, an iron layer was deposited on a copper substrate by means of electro-deposition method which could be irradiated by 29.5 MeV protons at 100 μA. No-carrier-added (n.c.a.) 55Co was separated from the iron target via an anion exchange column (Dowex 1 · 8). The achieved production yield was 31.25 MBq/μAh. Also, excitation functions for the 55Co radionuclide via natFe(p, xn)55Co, 56Fe(p, 2n)55Co and 54Fe(d, n)55Co reactions were calculated by TALYS-1.4 code and TENDL-2011 database and compared with previous published data.

This study is to investigate the issues on how to secure stability during the purification process for the production of recombinant protein A. The final recombinant protein A is produced by passing through the cation exchange column (SP) and the anion-exchange column (Q) during the production process, for which the samples produced by the step-by-step processes can be exposed to trouble in securing stable storage in case the next process cannot be taken within the proper time period. Accordingly, this study aims to evaluate the proper storage conditions and length of time when storing samples produced in the production process. That is, in this study, how to store fair samples, how long the storage period should be set up, and how to evaluate the security of its quality depending on time are dealt with. The items to be experimented with were enodotoxin, SDS-PAGE, HPLC purity and concentration. Experimental results showed that after passing the cation exchange column, when stored at 4 ℃C or room temperature, SDS-PAGE showed a major band, endotoxin is 5.0 Eu/mg or less, and concentration is on average of 8.21 to 8.24 mg/mL and RSD% 0.10~0.62%. In addition, HLPC purity showed somewhat stable results; at the HPLC purity 214 nm, the average is 99.24% to 99.37% and RSD% is 0.22~0.29%, while the average is 89.72% to 89.80% and RSD% 0.62~1.26% at 280 nm. On the contrary, after passing the anion exchange column, when stored at 4 oC or room temperature, SDS-PAGE revealed the major band, endotoxin is 0.5 Eu/mg or less, and concentration is on average of 5.59 mg/mL and RSD% 0.03~0.10%. when it comes to HLPC purity, the result showed that at the HPLC purity 214 nm, the average is 99.74% and RSD% is 0.10~0.11%, while the average is 96.16% to 96.85% and RSD% 0.72~1.13%. In conclusion, the stability of fair samples of recombinant protein A during the manufacturing process could be obtained without substance decomposition for 7~8 days at 4 ℃ or 20~21 days at room temperature.

In recent years, Cr speciation in dietary supplements has become decisive in the evaluation of their health risks. Despite being an beneficial micronutrient, Cr(III) can be toxic at living organisms at high concentrations, while Cr(VI) is known to be highly toxic and carcinogenic. The main objective of this work was to optimize an analytical methodology for the extraction and accurate quantification of Cr(III) and Cr(VI) in dietary supplements. The extraction of Cr species was carried out with 50mM EDTA solution on a hotplate under optimized conditions. Special attention was paid to bidirectional species transformations. No noticeable oxidation of Cr(III) into Cr(VI) was observed and the reduction to Cr(III) only occurred at very high Cr(VI) concentrations. Cr(III) as Cr(EDTA)(-) complex was chromatographically separated from Cr(VI), retained as CrO4(2-), on an anion exchange column coupled to inductively coupled plasma mass spectrometry (LC-ICP-MS). The limit of quantification (0.08µgg(-1)) was below the limit established for Cr enriched yeasts by the European Union. Eleven dietary supplements were analyzed and Cr(III) and Cr(VI) quantification was carried out by external calibration monitoring (52)Cr isotope and by speciated isotope dilution mass spectrometry (SIDMS) adding (50)Cr(III) and (53)Cr(VI) spikes. Total Cr was also quantified by ICP-MS and mass balance between total Cr and the sum of Cr(III) and Cr(VI) was achieved. In eight of the eleven tested supplements Cr(III) calculated amounts were higher than those indicated by the manufacturer, but only one of them exceeded the 250µgday(-1) recommended by World Health Organization (WHO). In contrast, it is worth noting that Cr(VI) amounts beyond the recommendations of the European Union for Cr enriched yeasts were found in five supplements. These results revealed that more accurate and rigorous quality assurance protocols should be applied to the testing of the final products, including the analysis of both Cr(III) and Cr(VI).

Particle accelerators have played a key role for the production of radioisotopes since the 1940s, and medical cyclotrons (11–20-MeV protons), in particular, are currently of central importance for the production of short-lived positron emitters for diagnostic applications in nuclear medicine. Commercially, cyclotrons in the 20–35-MeV proton energy range are used to produce a variety of gamma-emitting radioisotopes. In addition, many high-energy accelerators of several different types which accelerate primarily protons play a role in the production of medical radioisotopes, including those which have important roles in therapy. In this chapter, the basic fundamentals of accelerator production and yield calculations are discussed in addition to key therapeutic radioisotopes and comments on their applications as unsealed sources radiopharmaceuticals in nuclear medicine.

An ion chromatography method has been proposed for the determination of three common inorganic anions (chloride, nitrate and sulfate) in anionic surfactants using a single pump system. The new system consists of an ion exclusion column, a concentrator column, and an anion exchange column connected in series via two 6-ports valves in a Dionex ICS-2000 ion chromatograph. The valves were switched several times for removing surfactants, concentrating and separating the three anions. The chromatographic conditions were optimized. Detection limits (S/N = 3) were in the range of 0.10–0.68 μg/L. The relative standard deviations (RSDs) of peak area were less than 4.6%. The recoveries were in the range of 84.1–112.6%.

A method for the on-line absorptiometric determination of trace amounts of boron in steels was developed using an anion-exchange column presorbed with chromotropic acid. The on-line reaction and separation were achieved by controlling the pH in order to accelerate the complex formation in the column by 2.7 and to stabilize the complex at pH 8 for the selective elution of a 1:2 complex and its detection at 350 nm. The effects from the iron matrix in the sample were effectively removed by using EDTA as a masking agent; a low limit for boron detection (3σ) of 0.04 µg g(-1) was obtained.

A novel flow injection (FI) system is presented for simple, rapid and sensitive determination of cadmium at μg/g to sub-μg/g levels in iron and steel. The anion-exchange column incorporated in-line in a FI system was utilized as an integrated media for both the separation/preconcentration of cadmium from a large excess of iron matrix and the detection reaction of adsorbed cadmium with TMPyP (5,10,15,20-Tetrakis(N-methylpyridinium-4-yl)-21H,23H-porphine,tetrakis(p-toluene-sulfonate)) which was directly introduced into the column by switching its flow. The spectrophotometric detection at 512 nm for Cd complex with TMPyP was directly coupled in-line with such separation/preconcentration and reaction in this FI system which consists of a multi-position valve and a syringe pump and is operated automatically under the control by a personal computer. The variables relating to such reactions and manifold were studied in detail to establish the optimum conditions and manifold configulations. A linear calibration using a 400 μL sample injection was obtained for cadmium in the range of 0–20 μg/L (in the presence of 1.0×104 μg/mL iron matrix). The coefficient of variation for 10 μg/L cadmium was 1.8% (n=3), and the estimated limit of determination was 1 μg/L corresponding to 0.05 μg/g (5×10−6 mass%) in iron and steel sample. Only 14 min was required for an analytical procedure. Satisfactory recoveries of spiked cadmium were obtained to the solutions prepared by dissolving steel samples in acid mixtures.

In this paper, we describe the separation chemistry and electrodepositions conducted for the preparation of 241Am, 243Am and 233U targets used for cross-section measurements at DANCE. Thick, adherent deposits were prepared using molecular plating from isopropyl alcohol solutions. Improved yields and thicknesses were observed for 241Am electrodeposition after the material was purified using TRU resin from Eichrom. Similarly, 233U deposits were improved after purification with an anion exchange column in 9 M HBr followed by purification using UTEVA resin from Eichrom.

Samples of the vitreous were analysed in order to identify changes of soluble proteins in vitreo-retinal disease. The soluble proteins of the vitreous were separated on an anion exchange column (Mono-Q). The degree of neutral proteolytic activity in vitreous body was also measured. The vitreous from cataract cases without vitreoretinal disease was characterized by its low content of soluble proteins equivalent to about 1% of that of serum. Albumin and transferrin were the major identified components and their concentrations were approximately 0.85 and 0.03 g/l, respectively. In cases with vitreoretinal disease the vitreous showed changes of total soluble protein and the appearance of additional protein peaks. In patients with PVR the albumin concentration in the vitreous was found to be three times higher as compared to the control group consisting of patients with cataract. Neutral proteolytic activity in the vitreous was relatively low in both normal and pathological vitreous.

A practical analytical method was developed for the routine analysis of perchlorate in environmental and drinking-water samples by liquid chromatography-electrospray ionization mass spectrometry (LC/ESI-MS) using an anion exchange column. By using (18)O-enriched perchlorate as an internal standard, the limits of quantification of perchlorate determined by tenfold of the signal-to-noise ratio and tenfold of the standard deviation were 0.1 and 0.03 microg L(-1), respectively. The perchlorate concentrations in the raw and finished water samples from seven water purification plants were determined by LC/ESI-MS. Perchlorate was detected in 12 out of 13 samples, and the perchlorate concentrations in the samples were from 0.1 to 36.1 microg L(-1).

In this report, we describe how reactions of cell-free protein synthesis can be successfully conducted using plasmids prepared with regenerated anion-exchange columns. When washed, stripped, and equilibrated with appropriated buffers, regenerated columns were able to be used repeatedly to prepare plasmids with consistent yield and purity. The regenerated columns exhibited comparable performance to a fresh column with respect to the efficiency of protein synthesis using the plasmids prepared from them. Overall, we expect that the presented results will contribute significantly to economizing the technology of cell-free protein synthesis as a practical method for protein production in preparative scales.

A quantitative, reproducible, and efficient phytic acid assay procedure is needed to screen breeding populations and support genetic studies in soybeans [Glycine max (L.) Merr.]. The objective of this study was to modify the colorimetric Wade reagent method and compare the accuracy and applicability of this new method in determining seed phytic acid content in soybean with three well-established phytic acid assay methods: anion exchange column (AEC), high-performance liquid chromatography (HPLC), and 31 P nuclear magnetic resonance (NMR). The CV for repeated measurements of a low phytic acid soybean mutant, CX1834-1-6, ranged from 1.8 to 4.2% (n = 5), indicating the results were precise and reproducible. Phytic acid content of 42 soybean genotypes as determined by this method showed a correlation of 93.7 to 96.6% with the measurements by AEC, HPLC, and NMR. According to analysis of covariance, using inorganic P content as a predictor, phytic acid P content in a given sample analyzed by the four assay methods can be estimated with four linear regression models at the α = 0.01 level. Compared with HPLC, AEC, and 31 P NMR, this modified colorimetric method is simpler and less expensive for assaying a large number of samples, allowing its effective application in breeding and genetic studies of low phytic acid soybean.

Four unidentified acidic glycolipids (X3-X6) were isolated from the kidney of the Pacific salmon on an anion exchange column and by high performance liquid chromatography using a silica bead (Iatrobeads) column. Based on methylation analysis, chemical and enzymatic degradation, proton nuclear magnetic resonance spectroscopy and mass spectrometry, the glycon structure of X5 and X6 was identified as a unique disialosyl fucosyl-N-acetylgalactosaminyl ganglio-N-tetraose:Fucalpha3GalNAcbeta3Galbeta3GalNAcbeta4[NeuAcalpha8NeuAcalpha3] Galbeta4Glcbeta1Cer. NMR showed that X3 and X4 were analogues of X5 and X6 and contained O-acetyl groups on C4 of the outer N-acetylneuraminic acid, first disialosyl gangliosides containing 4-O-acetyl-N-acetylneuraminic acid. The ceramides of X3 and X5 contained predominantly C24: 1, and X4 and X6 contained saturated fatty acids (C14: 0, C16: 0 and C18: 0), whereas the long chain base was exclusively sphingenine. The concentrations of X3 and X4 were 0.13 and 0.16 nmol/g of kidney respectively and those of X5 and X6, were 0.07 nmol/g each.

(64)Cu is a useful radiotracer for positron emission tomography (PET) and a promising radiotherapy agent for the treatment of cancer. Recently, (64)Cu-labeled radiopharmaceuticals were reported to be useful for internal radiation therapy as well as PET monitoring of tumors.(64)Cu was produced at the Fukui Medical University's cyclotron using twelve MeV proton irradiation and the (64)Ni(p,n) (64)Cu nuclear reaction. A (64)Ni target was electroplated on a gold disk at a thickness of 50 to > 100 microm. Electroplating was performed at 2.5 V, at currents between 5-15 mA, and was completed in 12-24 hr. The (64)Ni target was bombarded with a 50 +/- 3 microA proton current. After bombardment, (64)Cu was separated from the (64)Ni target and other contaminants using an anion exchange column. Target (64)Ni was recovered and re-used. The yield of (64)Cu was 0.6 to > 3.0 mCi/microA*h, and averaged 1.983 mCi/microA*h. The radionuclidic purity of (64)Cu was over 99%. In this study, we obtained sufficient qualities and quantities of (64)Cu for therapeutic application and dose monitoring using PET using an ultra-small cyclotron.

A chemiluminescence (CL) flow-through sensor for ascorbic acid, based on permanganate electrostatically immobilized on an anion exchange column is described. Ascorbic acid can be measured by the CL reaction directly with the immobilized reagent. The sensors response was linear in the range 0.05–10 µg/mL with a detection limit of 5 ng/mL. The analysis could be performed within 30 s including sampling and washing, giving a throughout of about 120 samples per hour with RSD of 1.6% for 2 µg/mL ascorbic acid. The sensor was stable for over 500 determinations and has been successfully applied to the determination of ascorbic acid in tablets and vegetables.

Background: The small amount of allantoin present in human serum results from free radical (FR) action on urate and may provide a stable marker of free radical activity in vivo. We describe a gas chromatography–mass spectrometry (GC–MS) assay for serum allantoin and report a reference range in healthy individuals. Methods: Fasting blood samples were obtained from 134 healthy middle-aged volunteers (56 men, mean age 55, range 45–72; 78 women, mean age 55, range 50–72) Allantoin was assayed using 15N2 allantoin as an internal standard. After isolation from aqueous standards or serum by extraction onto an anion exchange column (AG-MP1), allantoin was derivatised with N-methyl-N-(tert-butyldimethylsilyl) trifluoroacetamide (MTBSTFA). Derivatives were injected onto an HP-1 column and analysed using a Mass Selective Detector with Single Ion Monitoring at 398 and 400 m/z. Results: The distribution of serum allantoin concentrations in men and women was non-Gaussian and log transformation was used for the analysis of data. Women (10.8±1.7 μmol/l (mean±S.D.)) had significantly lower serum allantoin levels than men (13.4±1.6 μmol/l, p=0.015). Reference ranges (95% CI) for middle-aged healthy subjects were 7.4–46.8 μmol/l (men) and 3.7–31.2 μmol/l (women). Conclusion: Gas chromatography–mass spectrometry provides a reliable and accurate method for the determination of serum allantoin.

A simple and specific liquid chromatographic method was developed for determination of clopidol in chicken tissues. Samples were extracted with acetonitrile. The extracts were cleaned up on an alumina column and an anion exchange column. The clopidol was separated on a column (30 cm × 3.9 mm) of μBondapak C18 (10 μm) by using acetonitrile–water (20 + 80, v/v) as mobile phase, and determined quantitatively at 270 nm. Recoveries were 86.0–97.6%, with relative standard deviations of 2.14–9.42% at 0.010–2.0 mg/kg from 4 spiked matrixes of chicken muscle, egg, liver, and kidney. The limit of detection was 0.005 mg/kg. Compared with the modified AOAC gas chromatographic method, the present method is simple and fast to operate. Its results are accurate and reliable, making it favorable for environmental protection and meeting requirements for human safety. Thus, it is suitable for routine analysis of large quantities of samples.

The white-rot fungus Marasmius quercophilus C30 is able to produce several laccases. The proportion of the enzymes produced depends on culture conditions. On malt medium, LAC1 was produced continuously over the 14 days of the cultivation period and was the only activity detectable. Copper increased total laccase activity by a factor 10 and induced the transient expression of one or more extra laccases in the culture medium. A combination of copper and p-hydroxybenzoic acid made it possible to extend the expression of induced laccase activities over the cultivation period and to reach a maximum activity 30 times higher than in non-induced culture. Extracellular laccases produced in this last condition were eluted as four peaks on an anion exchange column and were partially characterized.

A method is described for enhancing retention of arginine on a pellicular anion-exchange column. Arginine exhibits adjustable increases of retention time that are dependent on the acidity of standard or sample matrix. This effect is based on interactions of the protonated form of arginine with the residual cation-exchange groups on the core beads of pellicular particles. The relative magnitude of retention time shift of arginine is evaluated for identical concentrations of hydrochloric, sulfuric, and perchloric acids. Although the direct addition of acid is very effective in influencing the retention of arginine, it affects peak shapes and retention of other peaks in the chromatographic separation. The new technique—acid coinjection—achieves a similar retention enhancement for arginine with only a minimal effect on the rest of the separation. Detection limits, reproducibility results, and calibration data are presented for the chromatography of amino acids with acid coinjection. Improved resolution of arginine is demonstrated with chromatograms of soybean hydrolysate and cell culture samples.

A procedure for recovery of 65Zn from acidic Ga-rich solutions generated in 68Ge production process was developed and implemented in a “hot cell” environment. A one step simple separation was carried out on anion exchange column filled with Biorad AG1-X8 resin. With 65Zn recovery yields of more than 95% this procedure is currently routinely used at Brookhaven Linac Isotope Producer.

Rapid radiochemical neutron activation analysis (RNAA) procedures were developed and employed for the determination of 32 trace impurities in high purity aluminum thin foils. Anion exchange column chromatography was developed for the sequential group chemical separation of various elements which helped in reducing the spectral interferences and improving the sensitivity of the method. The procedure is simple and requires a very short time to separate the elements in three groups for radiometric assay. To determine very low contents of uranium and thorium,239Np and233Pa as activation products were separated using anion exchange and coprecipitation methods. The impurity contents were found to be low, therefore, their adverse effects on microelectronic devices would be negligible. Our data could partially be compared with the data reported in literature.

A comprehensive radiochemical procedure for the measurement of gamma/beta activity of131I has been standarized by optimizing different steps. The procedure is mainly divided into three stages, i.e., (1) concentration of iodine using anion exchange column followed by elution; (2) extraction of iodine into CCl4 and back extraction, and (3) precipitation as AgI. The percent chemical recovery and the lower limit of detection (LLD) were found to be 75.5±5% and 0.014 mBq/ml, respectively.

Using a biochemical approach we identified families of ubiquitin-activating and ubiquitin-conjugating enzymes in Chiamydomonas reinhardtii. The family of ubiquitin-activating enzymes, characterized by their ability to form thioesters with ubiquitin and eluting off a ubiquitin affinity column by ATP-depletion probably consists of at least four members. Whereas one of these enzymes is active under a broad range of pH values, thioesterof the other UBAs with ubiquitin is restricted to pH 7.5. Two ubiquitin-activating enzymes are metabolically phosphory-lated which is assumed to be an activity control mechanism. Most of the 7 ubiquitin-conjugating enzymes detected in this study were found to bind rather tightly to an anion exchange column, and eluted off the column at specific salt concentra tions. Two of the ubiquitin-conjugating enzymes described here did, however, not bind to this column. These enzymes can, as all other C. reinhardtii ubiquitin-conjugating enzymes, perform thioester formation with ubiquitin regardless of the source (plant/animal) of the ubiquitin-activating enzyme.