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  • Column liquid chromatography  (82)
  • pH  (44)
  • Springer  (126)
  • American Association for the Advancement of Science
  • Cambridge University Press
  • 1995-1999  (126)
  • 1975-1979
  • 1995  (126)
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Publisher
  • Springer  (126)
  • American Association for the Advancement of Science
  • Cambridge University Press
  • Wiley-Blackwell  (8)
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  • 1995-1999  (126)
  • 1975-1979
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  • 1
    ISSN: 1572-879X
    Keywords: metal-oxygen cluster compounds ; heteropoly acids ; stability ; pH ; aqueous solutions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The stabilities of the solid superacids H3Mo12O40, H3PW12O40, H4SiMo12O40 and H4SiW12O40 in aqueous solution have been measured at various values of pH by use of ion chromatographic analyses. The aforementioned acids are completely decomposed at values of pH, 4.0, 5.2, 7.0 and 11.0, respectively. The stabilities in aqueous solution with respect to pH follow the order H4SiW12O40 〉 H3PW12O40 〉 H4SiMo12O40 〉 H3PMo12O40.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Biology and fertility of soils 19 (1995), S. 197-202 
    ISSN: 1432-0789
    Keywords: Soil organic matter ; Cultivation ; CPMAS 13C-NMR ; Microbial biomass ; Substrate-induced respiration ; Alkylic carbon ; pH
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract To determine whether there is a relationship between the composition of soil organic matter and the activity of the soil microbial biomass, the composition of the organic matter in 12 typical arable soils in Northwest Germany was investigated by wet chemical analysis and CPMAS cross polarization magic angle spinning 13C-NMR spectroscopy. The data were correlated with the microbial biomass as estimated by substrate-induced respiration. A strong correlation between the microbial biomass and alkylic C compounds was observed (r=-0.960***). Recalcitrant substances were enriched in this fraction, which were classified as humic acids according to the wet chemical procedure. The microbial decomposition of these humic acids is probably retarded, due to their chemical structure and/or physical bonding, when the soil microbial biomass activity is limited.
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  • 3
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    Journal of comparative physiology 177 (1995), S. 503-510 
    ISSN: 1432-1351
    Keywords: pH ; Patch clamp ; Glutamate receptors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Proton-activated currents were examined in patch-clamp recordings from embryonic chick motoneurons. Rapid application of protons evoked a large inward current that peaked and then decayed, presumably due to channel inactivation. A pH shift from 7.4 to 7.1 was sufficient to evoke detectable currents. The shift from pH 7.4 required for half-maximal current amplitude (EC50) was to pH 6.8. In single-channel recordings, activation was achieved within 6 ms at pH 7. The average channel open time was 1.4 ms; the closed-state time constants were 1.0 and 6.2 ms. At pH 6.5, the single-channel conductance was 22 pS, and the reversal potential was similar to the calculated Na+ equilibrium potential. Current amplitude declined by 49% following addition of Ni2+ and increased by 58% as Ca2+ was lowered from 2 to 0.1 mM. Inactivation time constants ranged from 90 to 200 ms as pH varied from 6 to 7; these values did not depend on membrane potential. The reactivation time constant was 22 s. Proton- and glutamate-activated currents summated. Thus, transient decreases in extracellular pH can evoke large inward currents that decay rapidly and reactivate slowly. These currents may occur under pathological conditions that affect extracellular pH.
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  • 4
    ISSN: 1436-5073
    Keywords: polystyrene membranes ; fiber optics ; pH ; reflectance ; aminated polystyrene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract We have prepared toughened, porous, aminated polystyrene membranes that undergo an increase in reflectance as the pH increases from 6.8 to 8.0. Vinylbenzyl chloride (VBC) is copolymerized with divinylbenzene (DVB) in the presence of a toughening agent, Kraton G1652, a styrene-ethylene, butylene-styrene triblock copolymer, and a porogenic solvent, xylene/ dodecane. The optimum formulation for sensing is 2% DVB (mol DVB/mol VBC), 2% Kraton (g Kraton/g VBC) and 40% (v/v) 2∶1 xylene: dodecane. Benzoyl peroxide is used as the initiator. The components are partially polymerized at 85 °C to a viscosity of 600–800 centipoise. The polymerization is then stopped by reducing the temperature. A drop of the partially polymerized solution is confined between two microscope slides and the polymerization reaction is completed. The resulting membrane is then swollen in 1,4-dioxan and reacted with diethanolamine. These membranes have been incorporated into a pH sensor based on changes in reflected intensity measured through a bifurcated bundle of twenty unbuffered 50/55 core/cladding glass-on-glass optical fibers with numerical apertures of 0.57. The resulting sensor is stable and requires inexpensive optical components, a red-emitting LED as the source and a silicon photodiode as the detector.
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  • 5
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    Nutrient cycling in agroecosystems 44 (1995), S. 73-78 
    ISSN: 1573-0867
    Keywords: P precipitation ; precipitation kinetics ; P solubility diagram ; pH ; octacalcium phosphate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Orthophosphate (OP) is a major component of irrigation and nutrient solutions. Since OP precipitates may clog drippers, and deviations from intended OP concentrations may adversely affect plant development and yield, an understanding of the mechanisms controlling OP solubility in solutions of various ionic compositions, is essential. The objectives of this study were (i) to suggest guidelines for permitted OP additions to waters of various ionic compositions, so as to avoid OP crystallization and (ii) to predict the decrease in OP concentration as a function of time in supersaturated solutions. Five freshwater sources, used for irrigation in Israel, and representing extremes of pH and of Ca, HCO3 and SO4 concentrations, were tested. Solutions of three different initial OP concentrations (10, 30 and 90 mg Pl−1) and two pH values were prepared in 21 plastic bottles and kept in a dark room at 27 °C. Solution samples were withdrawn from the bottles at predetermined times, filtered and analysed for pH and total OP, Ca and HCO3 concentrations. In all the studied waters and for all initial OP levels the OP concentration (Cp) declined with time. The rate of decrease in Cp was proportional to the difference between the observed and equilibrium Cp values, with a specific rate constant for each water.The pH and the Ca2+ and HCO 3 − activities in solution were influenced by the initial Cp. The equilibrium Cp in all treatments was found to be controlled by octacalcium phosphate (OCP). Available chemical equilibria models allow to calculate the maximum level of OP that can be added to various waters before OCP precipitates, based on water pH and Ca, HCO3 and SO 4 2− concentrations.
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  • 6
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    Nutrient cycling in agroecosystems 44 (1995), S. 107-112 
    ISSN: 1573-0867
    Keywords: exchangeable Al ; exchangeable Ca ; ion pairs ; P sorption ; pH ; precipitation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract An earlier study of phosphate sorption by some savanna soils from Nigeria suggested that increased P sorption when pH was raised might be due to precipitation of exchangeable Al as amorphous polymeric Al species with increased sorption sites. But these savanna soils have Ca as the dominant cation in their exchange sites, and low exchangeable Al. The objective of this study was to determine the role played by Ca in pH-induced P sorption of three savanna soils under continuous cultivation. Phosphorus sorption increased when pH was raised from 4.5 to 7.0. Similarly, Ca retention increased with increasing pH. Regression of P sorption on Ca retention indicated a significant linear relationship in the three soils. Three possible mechanisms were proposed to explain the increasing P sorption with increasing pH: precipitation of Ca-phosphates, Ca-induced P sorption or co-adsorption of Ca and H2PO 4 − or HPO 4 2− as ion pairs or complexes. Available evidence suggests that all three mechanisms can operate together to enhance P retention as pH increases. The paper proposes that increased P sorption by savanna soils when pH is raised is likely to be related to the chemistry and retention of Ca rather than to hydrolytic reactions of Al.
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  • 7
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    Nutrient cycling in agroecosystems 45 (1995), S. 217-220 
    ISSN: 1573-0867
    Keywords: ammonia volatilization ; monoammonium phosphate ; pH ; urea triple superphosphate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Applying urea with acidic phosphate fertlizer increases urea fertilizer efficiency by reducing ammonia volatilization and toxicity to crop from urea hydrolysis. However, urea and triple superphosphate (TSP) are not recommended to be cogranulated because blends might become wet and sticky. Monoammonium phosphate (MAP) is a less acidic P source than TSP, but is compatible with urea. The objective of this study was to evaluate compound NP fertilizer products made from MAP and TSP combinations as P sources with urea. Fertilizer mixtures were pelletized from commercial urea, TSP and MAP with different N:P2O5 ratios and MAP/TSP combinations. Moisture changes during storage, pH of fertilizer solutions, and ammonia volatilization from surface applied fertilizer pellets were measured. Using MAP with TSP in urea-P mixtures reduced moisture increases during storage. Increasing MAP in urea-TSP-MAP combinations increased fertilizer solution pH by over 1 unit as the MAP/TSP-P2O5 ratio increased from 0/100 to 100/0. Adding MAP as 50% of P in urea-MAP-TSP mixtures at 3:1 and 1.5: (N:P2O5) ratios reduced ammonia loss from urea 50% to 60% compared to urea alone; and ammonia loss was similar to that of urea-TSP combinations. A urea-TSP-MAP fertilizer combination could make efficient use of urea-N by crops by reducing ammonia loss from urea hydrolysis.
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  • 8
    ISSN: 1432-1939
    Keywords: Detrital processing ; Exoenzymes ; ATP ; pH
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We examined microbial colonization, exoenzyme activity, and processing of leaves of yellow poplar (Liriodendron tulipifera), red maple (Acer rubrum), and white oak (Quercus alba) in three streams on the Allegheny Plateau of West Virginia, United States. Leaf packs were placed in streams that varied in their underlying bedrock geology, and therefore in their sensitivity to the high level of acidic precipitation that occurs in this region. The mean pH of the streams was 4.3 in the South Fork of Red Run (SFR), 6.2 in Wilson Hollow Run (WHR), and 7.7 in the North Fork of Hickman Slide Run (HSR). Through time, the patterns of microbial biomass and exoenzyme activity were generally similar among leaf species, but the magnitude of microbial biomass and exoenzyme activity differed among leaf species. Pectinase activity was greatest in HSR, the most alkaline stream, whereas the activity of exocellulase and xylanase was greatest in WHR and SFR, the intermediate and acidic streams. This variation in the activity of different exoenzymes was consistent with published pH optima for these exoenzymes. Variation in processing rates, both among leaf species and among streams, seems to be related to the level of microbial exoenzyme activity on the leaf detritus.
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  • 9
    ISSN: 1432-072X
    Keywords: Key words Spore activation ; Phycomyces ; Carbon source ; pH
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phycomyces blakesleeanus sporangiospores responded differently to activation by physical and chemical stimuli. Spores that were physically (heat shock) activated or chemically (ammonium acetate) activated germinated and grew at pH 4.5 with the hexoses glucose, fructose, galactose, and N-acetylglucosamine, and with glycerol and amino acids. Under these conditions, physically activated spores showed a lower, although significant growth with the hexoses fructose, galactose, N-acetylglucosamine and with glycerol. On the other hand, physically activated spores incubated at alkaline pH (pH 7.3) required glucose to germinate; a requirement not observed with chemically activated spores, which showed significant growth in the other hexoses tested. Both physically and chemically activated spores incubated at pH 7.3 were unable to germinate and grow with amino acids and glycerol. These results suggest that there are different targets for activation of the spores by physical and chemical treatments. The levels of the fermentative enzymes alcohol dehydrogenase and lactate dehydrogenase and of the oxidative enzyme NAD+-isocitrate dehydrogenase were higher in cells grown at pH 4.5 in medium containing glucose; however, alcohol dehydrogenase and lactate dehydrogenase appear not to be affected by a change in the pH of the growth medium.
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  • 10
    ISSN: 1573-2932
    Keywords: acidity ; aluminium ; metal cations ; pH ; soil organic carbon
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Surface waters in northern forest ecosystems receive a substantial amount of drainage water from superficial soil horizons enriched in organic matter (SOM). Chemical reactions in the interface between the soil solution andf organic colloides will therefore affect the surface water chemistry. The mobilization of total organic carbon (TOC) and pH was studied as a function of amounts of organically adsorbed Na, Ca and Al in two O and one A horizon, which differed in the likelihood of contributing to the chemistry in runoff, in a forested watershed in northern Sweden. The samples were hydrogen ion saturated, washed and titrated with NaOH, Ca(OH)2 and Al(OH)3 in a constant ionic medium of 0.01 M NaCl in order to give rise to a population of manipulated samples differing in the composition of adsorbed cations. The highly humified SOM accumulated in the Oh and Ah horizons of a Gleysol close to the draining stream was stabilized by flocculating Al (95% of adsorbed metal cations), which resulted in a low release of TOC. These horizons showed a high potential of organic carbon solubility when Al was changed for di- or monovalent cations. Calculations suggested that the release of TOC would increase more than ten times if Al was exchanged for Ca upon liming to pH 6.0. The pH values of all horizons were shown to be determined mainly by the composition of adsorbed mono-,di- and trivalent cations.
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  • 11
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    Water, air & soil pollution 85 (1995), S. 2185-2190 
    ISSN: 1573-2932
    Keywords: Cu ; Cu-Ni smelters ; Kola Peninsula ; Lapland ; Ni ; pH ; S ; snowpack
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Bulk snow samples were collected from the snowpack in open areas along two sampling lines running to the west from the Cu-Ni smelters at Nikel and Monchegorsk, NW Russia, during 1991–1993. The aim of the study was to estimate the area affected by sulphur and heavy metal deposition from the smelters. Snowpack quality was used as an indicator of deposition during winter time. The total sulphur, copper and nickel concentrations in the snowpack decreased significantly (p〈0.001) with increasing distance from the smelters along the sampling line running directly to the west from Monchegorsk. The deposition pattern was similar each winter during 1991–1993. The pH values did not correlate with the corresponding sulphur concentrations, and there was no decreasing pH gradient in the snowpack on moving towards Monchegorsk. The effects of sulphur emissions from Monchegorsk on snowpack chemistry were not detectable on the Finnish side of the border. The 3-year mean of the total sulphur concentration was 0.27 mg/kg, and of the pH values 4.92, along the sampling line running to the west of Monchegorsk. The total sulphur concentrations near the smelters (〈 20 km) varied between 0.37 and 0.95 mg/kg. The effect of the Cu-Ni smelters at Nikel on snowpack quality was not detectable in northern Finnish Lapland. The 3-year mean for total sulphur was 0.20 mg/kg and for pH 4.96 along the sampling line running to the west of Nikel.
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  • 12
    ISSN: 1573-2932
    Keywords: aluminium toxicity ; non-equilibrium chemistry ; pH ; stress ; apoptosis ; necrosis ; trout
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Mixing of acid river water containing aluminium (pH 5.1, Al 345 μg.l−1) with neutral water of a lake (pH 7.0, Al 73 μg.l−1) resulted in water (pH 6.4, Al 245 μg.l−1) with a pH (6.4) and Al concentration (245 μg.l−1) expected to have low toxicity to fish on the basis of current Al toxicity models. However, under semi-field conditions the freshly mixed water (a few sec. after mixing) proved to be highly toxic to brown trout. The fish were exposed to the water at different places along a 30 m channel. At the beginning of the channel acid and neutral water were continuously mixed; the mixed water left the channel after 340 sec. The cells of the gills showed a highly increased rate of cell death by apoptosis and necrosis. Intercellular spaces were enlarged, and many leucocytes penetrated in these spaces. Mucus release was stimulated to depletion. Plasma chloride levels were hardly affected. There was a clear gradient in the deleterious effects on the fish along the channel. The fish at the beginning of the channel (about 12 sec. after mixing of the water), were severely affected, whereas the fish kept at the end of the channel (340 sec. after mixing) were only mildly affected. In the natural situation fish will relatively quickly pass through a mixing zone. In our study we therefore focused on the effects on fish after a 60 min exposure to a mixing zone (5 sec after mixing), with subsequent recovery in a region downstream of the confluence and in neutral water with low Al. The recovery in the downstream area (at the end of the channel, i.e. 5 min after mixing) was clearly hampered when compared to the recovery in neutral water with low aluminium. Thus, a short exposure to the toxic mixing zone followed by a stay in water downstream of this zone, as may occur in nature, is detrimental to migrating trout. We conclude that freshly mixed acid and neutral water contain toxic components during the first seconds to minutes after mixing, that can not be explained by current models on aluminium toxicity.
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  • 13
    ISSN: 1573-2932
    Keywords: Scots pine ; Pinus sylvestris ; deposition ; element budget ; soil solution ; soil chemistry ; alkaline dust ; pH ; acidification ; sulfur release
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Since 1993 we are studying three Scots pine ecosystems along a deposition gradient in north-eastern Germany (formerly GDR). Dramatic reductions of pollutant emissions are reported for the period since 1989/90. S-deposition is high at the sites Roesa and Taura (25 kg S ha−1yr−1) compared to Neuglobsow. Inputs of basic cations, especially Ca, by alkaline dust immissions decrease in the order Roesa 〉 Taura 〉 Neuglobsow. The soil solution data show high concentrations of Ca and SO4 at Roesa decreasing drastically along the deposition gradient. The elevated pH values reflect the impact of alkaline dust deposition particularly in the organic surface layer at Roesa. The site Taura received less base cation deposition and is marked by the lowest pH values throughout the soil profile combined with increased Al concentrations in the solution of the mineral soil. Thus, the composition of the soil solutions clearly reflects the different deposition regimes of the past. The element budgets show that large amounts of base cations, sulfur, and, at Taura, also aluminum are actually released from the soils that were previously stored.
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  • 14
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    Water, air & soil pollution 85 (1995), S. 2125-2130 
    ISSN: 1573-2932
    Keywords: wet deposition ; sulphate ; pH ; snowfall ; Doppler weather radar ; short-range deposition modelling
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract The contribution to local wet deposition of emissions from a coal-fired power station at Inkoo on the south coast of Finland has been investigated during a wintertime precipitation event. Making use of intensive radiosonde and weather radar observations of meteorological factors, concentrations of sulphur in deposition due to plume washout were predicted by a short-range deposition model. The model used the scavenging coefficient to parametrize the wet removal of pollutants, and it took into account the wind drift of falling precipitation particles within the plume. The model predictions were then compared with the chemical analysis results from snowfall samples collected within 10 km of the power station during the experiment. The experiment was performed ahead of a deeply-occluded front during a period with strong advection of long-range transported pollutants. No reliable sign of the influence of the power station on the sulphate deposition could be identified. On the other hand, the deviations of acidity from the mean pH-value of 4.1 were concentrated in one sector near the expected area of deposited plume pollutants. If local emissions were responsible for these deviations, the explanation may lie in a slightly incorrectly estimated plume direction or the effects of alkaline fly ash. Nevertheless, definite conclusions cannot be drawn, because only a few collectors happened to be sited in the modelled sector of plume washout and none in its maximum area.
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  • 15
    ISSN: 1432-2048
    Keywords: Calcium ; Calmodulin antagonists ; pH ; SV-type channels (vacuole) ; Taproot, guard cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Slowly activating vacuolar (SV-type; Hedrich and Neher 1987, Nature 329: 833–835) ion channels provide the predominant membrane conductance of the vacuolar-lysosomal compartment of Vicia faba L. guard cells and sugar beet (Beta vulgaris L.) taproots. Applying the patch-clamp technique to isolated vacuoles of both tissues, the electrical and pharmacological properties of guard-cell SV-type currents were studied and compared to the sugar beet channel with regard to its modulation by cytoplasmic Ca2+ and pH. This outward rectifier of V. faba guard cells showed a half-maximum activation at 55–60 mV with an apparent gating charge equivalent of z ≈ 4. Studies on the single-channel and whole-vacuole level revealed an extremely high conductance of 280 pS for the guard-cell channels at a mean density of 0.37 μm-2 compared to taproots (120–140 pS at about 0.16 channels per μm2). Guard-cell SV-type channels are weakly selective for cations over anions and lack saturation at KC1 concentrations of up to 1 M. Since in the absence of physiological K+ concentrations, Ca2+ is the major permeable ion, relative changes in the amounts of the two ions might control the permeation process. In spite of their different origins and physiological functions, in guard cells and beet taproot cells, cytoplasmic Ca2+ and protons, both considered as candidates for intracellular signalling in plants, modulate the voltage dependence of SV-type channels. While the two effectors do not alter the single-channel conductance, they strongly interact with the voltage sensor. The calmodulin (CaM) antagonists N-(6-aminohexyl)-5-chloro-l-naphthalenesulfonamide hydrochloride (W-7), trifluoperazine (TFP) and calmidazolium hydrochloride (R 24571) effectively blocked the channel in an antagonist-specific manner. In agreement with the properties of a Ca2+-permeable channel, CaM could be involved in the modulation of the activation threshold of the SV-type channel. We therefore conclude that guard-cell SV-type channels, which might be responsible for the release of K+, Cl- and to a smaller extent Ca2+ during stomatal closure, could serve as an intracellular sensor for changes in cytosolic calcium (calcium-CaM) and pH.
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  • 16
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    The journal of membrane biology 145 (1995), S. 75-86 
    ISSN: 1432-1424
    Keywords: Amino acid transport ; ATP regulation ; Beta ; Mg2+ ; pH
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Macroscopic instantaneous and time-dependent currents have been measured in the vacuolar membrane of Beta vulgaris using a patch clamp configuration analogous to whole cell mode. At low cytosolic Ca2+ and in the absence of Mg2+, only an instantaneous current was observed. This current is carried predominantly by cations (PK∶PCl 7∶1, pna∶pcl 4∶1 and arginine is also conducted). The instantaneous current can be activated by ATP4− (e.g., ATP-activated mean K+ current density was −20 mA.m−2 at a membrane voltage of −20 mV) and by increasing cytosolic pH and Mg2+ (raising Mg2+ from 0 to 0.4 mm induced a mean current density increase of −7 mA.m−2 at −20 mV). Such current can be activated by simultaneous addition of putative in vivo concentrations of ATP4−/MgATP/Mg free 2+ (in the presence of bafilomycin to inhibit the vacuolar ATPase) and further modulated by cytosolic pH. With vacuolar K+ concentration greater than that of the cytosol, activation of the instantaneous current would mediate vacuolar K+ release over the range of physiological membrane voltage. It is argued that the ATP4−-activated current, in addition to acting as a K+ mobilization pathway, could provide a counter-ion (shunt) conductance, allowing the two electrogenic H+ pumps which reside in the vacuolar membrane to acidify the vacuolar lumen. A separate time-dependent current, which was not observed at low Ca2+ concentrations (less than 500 nm) could also be elicited by addition of Mg2+ at the cytoplasmic membrane face. This current was stimulated by increasing cytoplasmic pH.
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  • 17
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    The journal of membrane biology 147 (1995), S. 255-262 
    ISSN: 1432-1424
    Keywords: Labyrinth ; Slowly activating K+ channel ; IsK channel ; MinK channel ; pH
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The pH-sensitivity of transepithelial K+ transport was studied in vitro in isolated vestibular dark cell epithelium from the gerbil ampulla. The cytosolic pH (pH iwas measured microfluorometrically with the pH-sensitive dye 2′,7′-bicarboxyethyl-5(6)-carboxyfluorescein (BCECF) and the equivalent short-circuit current (I sc), which is a measure for transepithelial K+ secretion, was calculated from measurements of the transepithelial voltage (V t)and the transepithelial resistance (R t) in a micro-Ussing chamber. All experiments were conducted in virtually HCO 3 − -free solutions. Under control conditions, pH iwas 7.01±0.04 (n=18), V twas 9.1±0.5 mV, R t16.7±0.09 Ωcm2, and I sc was 587±30 μA/cm2 (n=49). Addition of 20 mm propionate− caused a biphasic effect involving an initial acidification of pH i, increase in V tand I sc and decrease in R tand a subsequent alkalinization of pH i, decrease of V tand increase of R t. Removal of propionate− caused a transient effect involving an alkalinization of pH i, a decrease of V tand I sc and an increase in R t. pH iin the presence of propionate− exceeded pH iunder control conditions. Effects of propionate − on V t, R tand I sc were significantly larger when propionate− was applied to the basolateral side rather than to the apical side of the epithelium. The pH i-sensitivityof I sc between pH 6.8 and 7.5 was −1089 μA/(cm2 · pH-unit) suggesting that K+ secretion ceases at about pH i7.6. Acidification of the extracellular pH (pH o)caused an increase of V tand I sc and a decrease of R tmost likely due to acidification of pH i. Effects were significantly larger when the extracellular acidification was applied to the basolateral side rather than to the apical side of the epithelium. The pH osensitivity of I sc between pH 7.4 and 6.4 was −155 μA/(cm2 · pH unit). These results demonstrate that transepithelial K+ transport is sensitive to pH iand pH oand that vestibular dark cells contain propionate− uptake mechanism. Further, the data suggest that cytosolic acidification activates and that cytosolic alkalinization inactivates the slowly activating K+ channel (I sK)in the apical membrane. Whether the effect of pH ion the I sK channel is a direct or indirect effect remains to be determined.
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  • 18
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    The journal of membrane biology 148 (1995), S. 1-11 
    ISSN: 1432-1424
    Keywords: Toad skin ; pH ; Ion conductance ; Voltage dependence ; Chloride conductance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The present study focuses on two closely related topics on ion conductance in toad skins: (i) the interaction of apical protons with the apical voltage-dependent Cl−-activated channels of the mitochondriarich cells, and (ii) the description and characterization of a novel subject, a voltage-dependent H+-activated conductance. The Cl− conductance (G Cl) is activated by tissue hyperpolarization (which leads to apical membrane depolarization) and the presence of Cl− ions in the apical solution. Increasing apical proton concentration (from pH 8 to pH 4) impairs the process of activation of the Cl− conductive pathway, slowing the kinetics of I t activation and reducing the steady-stage values of G t and I t . This effect is markedly voltage-dependent since no effect is seen at V t =−100 mv and is fully present at −50 mV. The voltage-dependence of the pH effect suggests that the critical protonation sites of the apical Cl− channels are not freely exposed to the apical solution but dwell within the membrane electric field. An also coherent interpretation is that titration of apical proton binding sites affects the gating of the voltage-dependent Cl− channels, shifting the conductance-vs.-voltage curve to more negative clamping potentials. Tissue conductance in the absence of apical Cl− ions can be importantly affected by the pH of the apical solution (pH a ), the effect being markedly dependent on the clamping potential. Generally speaking, the effect of rising apical proton concentration can be conspicuous at negative clamping potentials, while at positive potentials changes in tissue conductance were never observed. For a clamping potential of −100 mV, a turning point somewhere between pH a =4 and pH a =3 was observed. Apical acidification to pH 4 has no effect upon tissue conductance while apical acidification to pH 3 leads to a marked, slow and reversible increase of tissue conductance. A striking similitude exists between the voltage-dependent Cl−-gated conductance and the voltage-dependent proton-gated conductance regarding: (i) slow time courses of activation and deactivation, (ii) requirement for a negative clamping potential and the presence of a specific ion species in the apical solution for activation to take place, (iv) instantaneous ohmic behavior, and (v) steady-state rectification. However, so far the results do not permit one to conclude definitely that the voltage-dependent Cl−-gated conductance and the voltage-dependent proton-gated conductance share a common pathway.
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  • 19
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    The journal of membrane biology 147 (1995), S. 207-215 
    ISSN: 1432-1424
    Keywords: Rana esculenta oocytes ; H+ current ; pH ; Voltage clamp
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Voltage clamp technique was used to study macroscopic ionic currents in Rana esculenta oocytes. Depolarization steps led to the activation of a single type of outward current (I out) when contaminant potassium and calcium-dependent chloride currents were pharmacologically inhibited. The voltage threshold of I out activation was 10 mV and this current, which did not inactivate, presented a deactivation the time constant of 73±21 msec (n=26) corresponding to a membrane voltage of −60 mV. Its reversal potential (E rev) was dependent on the magnitude of the depolarization and also on pulse duration. These changes in E rev were thought to reflect intracellular ion depletion occurring during activation of the remaining outward current. Furthermore, the activation threshold of I out was clearly affected by modifications in extracellular and intracellular H+ concentrations. Indeed, intracellular alkalinization (evoked by external application of ammonium chloride) or extracellular acidification induced a rightward shift in the activation threshold while intracellular acidification (evoked by external application of sodium acetate) or extracellular alkalinization shifted this threshold toward a more negative value. Lastly, I out was dramatically reduced by divalent cations such as Cd2+, Ni2+ or Zn2+ and was strongly decreased by 4 Aminopyridine (4-AP), wellknown H+ current antagonists already described in many cell types. Therefore, it was suggested that the outward current was prominently carried by H+ ions, which may play a key role in the regulation of intracellular pH and subsequent pH dependent processes in Rana oocyte.
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  • 20
    ISSN: 1432-1424
    Keywords: Membrane potential ; Conductances ; pH ; Acetate ; NH4C1
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Intracellular pH (pH i ), membrane potential (V m ) and membrane conductance (G m ) in fused proximal tubular cells of the frog kidney, were determined at three extracellular pH (pH o ) values, 7.5, 8.5 and 6.5. Imposed changes of pH o by ±1 pH unit induced parallel but smaller shifts of pH i . The alkaline milieu hyperpolarized the cells and increased G m , whereas the acid milieu depolarized and lowered G m . We subsequently introduced a weak acid and its conjugate base (acetic acid/acetate), or a weak base and its conjugate acid (NH3/NH 4 + ), at pH o 7.5, 8.5 and 6.5 to shift pH i -without altering pH o , or to shift pH i against imposed changes of pH o . From these experiments, we observed that under some circumstances V m varied with pH o but without G m or pH i changes, whereas under other circumstances changes of G m occurred during alterations of pH i while pH o and V m remained unaltered. At pH i ≈ 6.5 associated with V m ≈ −10 mV, G m dramatically increased to quasi-infinite values. This increase was not an artifact since G m returned to its control value following recovery to the control solution or in the presence of hyperosmotic solution. In conclusion, we demonstrate a differential regulation whereby V m and G m are controlled by pH o and pH i : pH o modulates mainly V m , and pH i modulates chiefly G m . Furthermore, at pH i ≈ 6.5 and V m ≈ −10 mV, our data reveal a large G m that tends towards infinite values in a reversible fashion.
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  • 21
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    The journal of membrane biology 147 (1995), S. 121-136 
    ISSN: 1432-1424
    Keywords: Chloride channel ; Cardiac sarcoplasmic reticulum ; Planar lipid bilayer ; Ion selectivity ; Voltage ; Block ; pH
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract We have characterized a voltage-sensitive chloride channel from cardiac sarcoplasmic reticulum (SR) following reconstitution of porcine heart SR into planar lipid bilayers. In 250 mm KCl, the channel had a main conductance level of 130 pS and exhibited two substrates of 61 and 154 pS. The channel was very selective for Cl− over K+ or Na+ ( $$P_{{\text{K}}^{\text{ + }} } /P_{{\text{Cl}}^{\text{ - }} } = 0.012$$ and $$P_{{\text{Na}}^{\text{ + }} } /P_{{\text{Cl}}^{\text{ - }} } \sim 0.040$$ ). It was permeable to several anions and displayed the following sequence of anion permeability: SCN− 〉 I− 〉 NO 3 − ∼ Br− 〉 Cl− 〉 f− 〉 HCOO−. Single-channel conductance saturated with increasing Cl− concentrations (K m= 900 mm and γmax = 488 pS). Channel activity was voltage dependent, with an open probability ranging from ∼1.0 around 0 mV to ∼0.5 at +80 mV. From −20 to +80 mV, channel gating was time-independent. However, at voltages below −40 mV the channel entered a long-lasting closed state. Mean open times varied with voltage, from ∼340 msec at −20 mV to ∼6 msec at +80 mV, whereas closed times were unaffected. The channel was not Ca2+-dependent. Channel activity was blocked by disulfonic stilbenes, arylaminobenzoates, zinc, and cadmium. Single-channel conductance was sensitive to trans pH, ranging from ∼190 pS at pH 5.5 to ∼60 pS at pH 9.0. These characteristics are different from those previously described for Cl− channels from skeletal or cardiac muscle SR.
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  • 22
    ISSN: 1432-1424
    Keywords: Na+/H+ exchange ; H+-ATPase ; Proximal tubules ; Kidney ; Acid-base ; pH
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract This study examines the effects of acute in vitro acid-base disorders on Na+/H+ and H+-ATPase transporters in rabbit kidney proximal tubules (PT). PT suspensions were incubated in solutions with varying acid base conditions for 45 min and utilized for brush border membrane (BBM) vesicles preparation. BBM vesicles were studied for Na+/H+ exchange activity (assayed by 22Na+ influx) or abundance (using NHE-3 specific antibody) and H+-ATPase transporter abundance (using antibody against the 31 kDa subunit). The Na+/ H+ exchanger activity increased by 55% in metabolic acidosis (pH 6.5, HCO 3 − 3 mm) and decreased by 41% in metabolic alkalosis (pH 8.0, HCO 3 − 90 mm). The abundance of NHE-3 remained constant in acidic, control, and alkalotic groups. H+-ATPase abundance, however, decreased in metabolic acidosis and increased in metabolic alkalosis by 57% and 42%, respectively. In PT suspensions incubated in isohydric conditions (pH 7.4), Na+/H+ exchanger activity increased by 29% in high HCO 3 − group (HCO 3 − 96 mm) and decreased by 16% in the low HCO 3 − groups (HCO 3 − 7mm. The NHE-3 abundance remained constant in high, normal, and low [HCO 3 − ] tubules. The abundance of H+-ATPase, however, increased by 82% in high [HCO 3 − ] and decreased by 77% in the low [HCO 3 − ] tubules. In PT suspensions incubated in varying pCO2 and constant [HCO 3 − ], Na+/H+ exchanger activity increased by 35% in high pCO2 (20% pCO2, respiratory acidosis) and decreased by 32% in low pCO2 (1.5% pCO2, respiratory alkalosis) tubules. The NHE-3 abundance remained unchanged in high, normal, and low pCO2 tubules. However, the H+-ATPase abundance increased by 74% in high pCO2 and decreased by 69% in low pCO2 tubules. The results of these studies suggest that the luminal Na+/H+ exchanger is predominantly regulated by pH whereas H+-ATPase is mainly regulated by [HCO 3 − ] and/ or pCO2. They further suggest that the adaptive changes in H+-ATPase transporter are likely mediated via endocytic/exocytic pathway whereas the adaptive changes in Na+/H+ exchanger are via the nonendocytic/exocytic pathway. The excellent technical assistance of Yollanda J. Hattabaugh, Gwen L. Bizal, and L. Yang is greatly appreciated. Portions of these studies were presented at the annual meeting of the American Society of Nephrology, Boston, MA, November 1993, and published in abstract form (J.Am.Soc.Neph. 4:840A, 1993)
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  • 23
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    The journal of membrane biology 148 (1995), S. 83-90 
    ISSN: 1432-1424
    Keywords: Cation channel ; pH ; Nucleotides
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract We investigated the effects of internal pH on Ca-activated, nucleotide-inhibited nonselective cation channels in the basolateral membranes of mouse collecting tubules, using the inside-out variant of the patch clamp technique. pH modulated the channel open probability (P o ), giving a bell-shaped curve peaking at pH 6.8/7.0: P o at pH 6.0 was 11±6% of P o at pH 7.2 and 32 ±7% at pH 8.0. The open and closed time distributions, best fitted to the sum of two exponentials, were differently sensitive to acid and alkaline conditions. Low pH reduced the short and long open times to 38 and 24% of their pH 7.2 values, while high pH produced a 4-fold increase in the long closed time. As previously reported, 4-acetamido-4′-isothiocyanatostilbene-2,2′-disulfonic acid (SITS) induced a quasi-permanent opening of the channel. The inhibition of the channel produced by high pH disappeared in the presence of SITS, while the inhibition produced by low pH was unaffected. These results suggest that the pH dependence of the channel is due to two separate mechanisms. pH was without effect on the ATP-evoked inhibition of the channel, while high pH profoundly reduced the steepness of the AMP inhibition curve, without altering the half-maximal inhibitory AMP concentration.
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  • 24
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    Mycoscience 36 (1995), S. 243-246 
    ISSN: 1618-2545
    Keywords: city waste ; fungal growth ; pH ; temperature
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    Topics: Biology
    Notes: Abstract Relationships between the growth of certain fungi isolated from city waste and pH and temperature were examined by two methods. The tested isolates showed their maximum growth and sporulation at different pHs while temperature requirements were the same (28°C), except forHumicola grisea (43°C).Cladosporium herbarum andH. grisea showed double pH optima. The ranges of pH and temperature for sporulation were more limited than those for the vegetative growth. Although all the tested isolates showed wide tolerances to pH and temperature, the degree of tolerance varied with the isolates. A considerable change from the initial pH of the liquid medium was noted at the end of the experiment.
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  • 25
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    Chromatographia 40 (1995), S. 99-101 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; High temperature open-tubular columns ; Solvent make-up ; Acetophenone oxidation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Acetophnone, a substance stable at room temperature, is readily oxidizes in a high temperature open tubular liquid chromatographic system by the residual oxygen in the mobile phase. The oxidation product is less UV absorptive and detection sensitivity decreases greatly. To prevent the oxidation, through degassing of the mobile phase is necessary.
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  • 26
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Endotoxins ; Lipopolysaccharide ; Aminated poly(γ-methyl L-glutamate) ; Bovine serum albumin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A method is described for the selective removal of endotoxins from various protein solutions using columns packed with aminated poly (γ-methyl L-glutamate) (PMLG-NH2) spheres. The PMLG-NH2 adsorbents showed a high adsorbing activity for endotoxins which had an ionic strength of μ=0.05–1.0 and pH 5.0–9.0. The endotoxin-adsorbing capacity per millitre of the wet adsorbent increased from 0.40 to 1.35 mg (E. coli O111: B4 LPS) at μ=0.2 and pH 7.0 while the aminogroup content of the adsorbent increased from 0.8 to 3.5 meq g−1. The PMLG-NH2-3.5 has an amino-group content of 3.5 meq g−1. This column packing selectively adsorbed endotoxins, without loss of the protein, from a γ-globulin or cytochromec solution which contained endotoxins at μ=0.05 and pH 7.0. On the other hand, when bovine serum albumin (BSA) was present in solution with endotoxins, both the endotoxins and the BSA were adsorbed by the column. The BSA-adsorbing activity increased with increasing amino-group content of the adsorbent. However, this undesirable adsorption was suppressed with increasing ionic strength of the buffer. As a result, when the packing which had an amino-group content of 1.5 meg g−1 was used in conditions of μ=0.2 and pH 7.0, the endotoxins were removed from a BSA-containing solution without affecting the recovery of the BSA.
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  • 27
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    Chromatographia 40 (1995), S. 51-57 
    ISSN: 1612-1112
    Keywords: Column electrophoresis ; Column liquid chromatography ; Phenolic lignin degradation compounds ; Sugar degradation products ; Quantitative analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A method for the quantitative analysis of phenolic lignin degradation products by capillary electrophoresis (CE) with on-column UV detection has been developed. The liquid biomass solutions contain low molecular hemicellulosic sugars and phenolic lignin degradation products with various degrees of polymerization. Special attention has been paid to the monomeric phenolic components of lignin degradation fragments, e.g. derivatives of phenolic acids, aldehydes, and alcohols. Uncoated fused silica capillaries and borate-phosphate buffer systems at moderate pH conditions were used in order to separate the compounds of interest. To provide validation of the method, the same samples were analyzed independently by HPLC using gradient elution on a RP-C18 column. As sugar degradation fragment, furan-2-carboxylic acid was detected.
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  • 28
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    Chromatographia 40 (1995), S. 85-90 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Phenolic compounds ; Ion-pair solid phase extraction ; On-line trace enrichment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Two packing materials, C18 and PLRP-S, are studied for on-line trace enrichment of phenolic compounds in water. Various precolumns of different internal diameter are also tested and the addition of an ion-pair reagent to increase retention and thus, breakthrough volumes of phenolic compounds, is studied. Best results are obtained when a PLRP-S precolumn is coupled on-line with a C18 analytical column and DAD detector. Addition of TBA considerably increases breakthrough volumes. In contrast, when a C18 precolumn is used, breakthrough volumes are lower and it is impossible to determine TCP and PCP, under the experimental conditions used, because of interference of other nonpolar compounds in the samples. The performance of the system is evaluated with river and tap water and the preconcentration of 10 ml of sample in a PLRP-S precolumn involves a linear range between 1 μg 1−1 and 20 μl−1 and limits of determination between 0.5 μg l−1 and 1 μg l−1 are obtained.
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  • 29
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    Chromatographia 40 (1995), S. 631-637 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Trace analysis ; Pesticides in tap water ; Aromatic sulphonates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Methodology and instrumentation are described which allow chromatographic screening of water samples under substantially simplified conditions and at reduced cost. A single column is used to accomplish sample extraction, trace enrichment, and separation. The performance of such a system is demonstrated, and the results compared with conventionally used concepts.
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  • 30
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Transchelation ; Zinc pyrithione and related compounds ; Antidandruff formulations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary An analytical procedure has been developed for the determination of zinc pyrithione in antidandruff formulations. Zinc pyrithione was converted into a stable copper complex and then analysed by reversed-phase HPLC. The proposed method allows the separation of the analyte from related pyrithiones and therefore is able to verify the compliance of cosmetic preparations with current legislation.
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  • 31
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Benzodiazepines ; Solvent effects ; ACN clusters
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Two different methods were used to determine the separation factor α at different temperatures and the Gibbs-Helmholtz parameters (Δ(ΔH), Δ(ΔS)) of two adjacent benzodiazepines on a chromatogram were obtained from plots of lnα versus 1/T. We first studied each factor (fraction of water ϕ in the ACN/water mixture and column temperatureT), which controls the retention mechanism, and then we examined the simultaneous variation of all these factors. The changes in Δ(ΔH) and Δ(ΔS) in relation to a volume fraction of water ϕ in an ACN/water mixture were examined. In the ACN/water system, Δ(ΔH) was fairly constant in the acetonitrile region of ϕ≤0.52 and appears to be a roughly linear function of ϕ for ϕ≥0.52. In this system Δ(ΔS) is approximately a parabolic function of ϕ with an optimum at ϕ≅0.52. The retention mechanism of ten benzodiazepines was found to be significantly different in the methanol/water and ACN/water mixtures. The separation optimization of these ten benzodiazepines was then considered. A fraction of water of 0.43 in the ACN/water mixture and a column temperature of 44°C gave the most efficient separation conditions in the ACN/water mixture.
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  • 32
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Polyurethane degradation products ; 2,4- and 2,6-toluendiamine ; Urine analysis ; Breast implants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A reversed-phase HPLC method has been developed for the urinary determination of mutagenic 2,4- and 2,6-toluendiamines. These amines are degradation products of polyurethane, a material used to cover textured silicone breast implants. FMOC-Cl was used as fluorescent derivatising agent in order to obtain a limit of detection of 15 ng/ml in urine. Pre-treatment of urine samples was by liquid/liquid extraction and urine specimens of patients after surgury were analysed.
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  • 33
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    Chromatographia 41 (1995), S. 657-660 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Polyphenols ; Flavonoids ; Phenolic acids and aldehydes ; Eucalyptus spp.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Standards of the polyphenols occurring in wood, bark and leaf extracts ofEucalyptus spp. (i.e. flavonoids and phenolic acids and aldehydes) have been analyzed by HPLC using reversed phase columns, gradient elution and diode-array detection. The conditions used are reported.
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  • 34
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    Chromatographia 40 (1995), S. 657-661 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Reversed phases ; Polyencapsulated silica ; Polyacrylates
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    Topics: Chemistry and Pharmacology
    Notes: Summary By copolymerization of silica modified with vinyl groups and acrylic acid esters in suspension highly efficient, polymeric-encapsulated stationary-phases can be prepared. The suspending liquid has to be selected such that the monomers are at least partially adsorbed on the surface and the polymer formed does not precipitate. Under these conditions stationary phases capable of speedy mass transfer can be prepared, where the reduced plate heights are between 2 and 3. The silica surface shielding is optimized so that basic and acidic solutes elute with symmetrical peak.
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  • 35
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Chiral resolution of amino acids ; Pre-column derivatization ; Fluorescent chiral tagging reagents ; Stereochemical purity
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    Topics: Chemistry and Pharmacology
    Notes: Summary Derivatization of amino acid enantiomers with fluorescent chiral Edman-type reagents, 4-(3-isothiocyanatopyrrolidin-1-yl)-7-nitro-2,1,3-benzoxadiazole [R(−)- and S(+)-NBD-PyNCS] and 4-(3-isothiocyanatopyrrolidin-1-yl)-7-(N,N-dimethylaminosulfonyl)-2,1,3-benzoxadiazole [R(−)-and S(+)-DBD-PyNCS], yields corresponding diastereomers separable by reversed-phase HPLC on normal achiral columns. The resolved diastereomers were detected fluorometrically at 530 nm with excitation at 490 nm for the NBD-PyNCS derivatives and at 560 nm with excitation at 450 nm for those derived from DBD-PyNCS reagents. This HPLC-derivatization method was used for evaluation of stereochemical purity for some synthetic commercial peptides. The enantioanalysis was reliable down to 0.05% racemization of the amino acid residues and a quantity of 100 μg peptide sample was usually enough for the analysis. Two acid hydrolysis methods, i.e. the standard procedure with constant-boiling hydrochloric acid (HCl) and a rapid vapor-phase procedure with HCl-trifluoroacetic acid (TFA) mixture, were compared. The later was found to be unsuitable owing to increased racemization of the amino acid residues during the hydrolysis. Judging from the results obtained for proline and leucine residues, most of the tested peptides including biologically active peptides, such as neurotensin, [d-Ala2,d-Leu5]-enkepharin and morphine tolerance peptide, possessed stereochemical purities higher than 98%. Influence of structural features of the peptides on the racemization of the amino acid residues was found to be significant.
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  • 36
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Mass spectrometric detection ; Fast atom bombardment ionization ; Serum bile acid ; Rat bile
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary In order to analyse bile acids in biological matrices, e.g. rat bile and human serum, high performance liquid chromatography (HPLC) was coupled to continuous-flow fast atom bombardment mass spectrometry (CFFAB-MS). A gradient elution system which had already proved to be well suited for the quantitative determination of conjugated bile acids in bile was modified to allow HPLC-CFFAB-MS-coupling. Due to the sensitivity of this coupling method it is possible to obtain more information about the biliary bile acid pattern and species-specific secondary bile acids. Furthermore, we were able to identify obviously unknown bile acid species in rat bile which most likely classify as mono-oxo and di-oxo-taurocholates (MW 513 Da, 511 Da) and mono-oxo-glycocholates (MW 463 Da). In the present study we show that using this system it is possible to determine both conjugated and unconjugated as well as sulfated bile acids, without time consuming group separation and derivatization, from rat bile and human serum. In addition, it is suggested that the method presented here should be considered for use in routine analysis.
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  • 37
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Fluorescence enhancement ; Bovine serum albumin ; Dansylamino acids
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    Topics: Chemistry and Pharmacology
    Notes: Summary The effects of acetonitrile and bovine serum albumin (BSA) concentrations on the signal intensity and retention behavior of dansylamino acids have been examined by using γ-cyclodextrin-bonded silica gel as the stationary phase in microcolumn liquid chromatography. Fluorescence intensities of dansylamino acids were enhanced by BSA as a mobile phase additive, e.g., detection limits of dansyl derivatives of L-Ala and L-Phe were improved by a factor of 12–18.
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  • 38
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Micellar liquid phases ; Octanol-water partition coefficient ; Benzene and naphthalene derivatives
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary It is shown that the logarithm of the capacity factor (log k′) for fifteen benzene and naphthalene derivatives in micellar liquid chromatography with forty nine different mobile phases generally correlates better with the logarithm of the octanol-water partition coefficient (log Pow) than the capacity factor (k′). Optimum conditions are established to obtain the best linear correlations of log k′-log Pow.
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  • 39
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Photolysis ; Electrochemical detection ; Drugs with chlorinated aromatic rings
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Pharmaceuticals containing a thiazide ring or chlorinated aromatic ring were investigated with respect to enhanced selectivity in determination. Oxidative electrochemical detection coupled with HPLC was used to study the influence of the pH of the mobile phase under conditions of photolysis. To cover a pH range 3.9–12, the employment of a polymer column stable in alkaline media was necessary. The method offers the great advantage of derivatization without chemicals at low operating potentials, thereby providing high selectivity.
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  • 40
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    Chromatographia 40 (1995), S. 296-302 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Structural isomer separation ; Cyclodextrin ; Polymers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Several β-cyclodextrin (CD)-bonded stationary phases for high performance liquid chromatography (HPLC) were prepared, based on silica beads coated with a poly(alkylamine), [poly(ethyleneimine)(PEI)]. In order to obtain stationary phases with a high content of CD and maximum accessibility of the CD cavity, the functionalization was carried out after the coating, using the mono-tosyl derivative of CD as the intermediate. The ability of these supports to separate ortho, meta and para isomers of some disubstituted benzene derivatives was examined. The contribution of the amino groups of the polymer and of the CD cavity to the separation process is discussed. The resolution is mainly based on the difference in the stability of the complexes of the various isomers with CD. The influence of the amount of CD on the height of the theoretical plate is also studied.
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  • 41
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Reversed phase ; Polystyrene ; Pellicular ; Ultra high molecular mass
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Ultra high molecular mass (〉 7 million dalton) polystyrenes are prone to degradation in size exclusion chromatography. In gradient elution reversed phase HPLC they do not elute visibly on small particle size porous supports. However, large diameter C18 pellicular particles were successfully employed for reversed phase study of ultra high molecular mass (15 million dalton) polystyrenes without polymer degradation during elution. Although retention for the lower molecular mass polystyrenes was lower than on small diameter porous particles, the medium high molecular mass polystyrenes (0.5–1 million dalton) showed similar retention. The addition of small diameter porous particles in small quantities, to the large diameter pellicular particles, increased the amount of retention of the low molecular mass polystyrenes without affecting the higher molecular mass polystyrenes.
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  • 42
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Methyl ethyl ketone in urine ; Derivatization ; 3-Methyl-2-benzothiazolinone hydrazone ; Comparison with GC-MS
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A HPLC method for the determination of methyl ethyl ketone (MEK) in urine after derivatization with 3-methyl-2-benzothiazolinone hydrazone is proposed. The calibration curve for the ketone was linear, ranging between 0.23–10 mg/L, with a detection limit of 0.025 mg/L. The results were compared to those obtained by GC-MS, coupled to the headspace technique. MEK derivatization and the derivative purification processes were verified with respect to the main variables such as reaction temperature, reagent concentration, probable interferences and enrichment phase. The method is simple and reliable and shows a good sensitivity.
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  • 43
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    Chromatographia 40 (1995), S. 382-386 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Sulfonamides in foodstuffs ; Honey ; Milk ; Eggs
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A procedure for the simultaneous determination of several sulfonamides in different foods, such as honey, milk and eggs is proposed. The analysis is carried out using reversed phase liquid chromatography with spectrophotometric detection. Optimization of the mobile phase led to good separation and a short analysis time when an initial isocratic step with a 3∶97 acetonitrile: water mixture was used for 5 minutes, followed by a linear gradient up to a 40∶60 mixture over 15 min. The proposed method is suitable for routine quality control analysis to ensure the absence of sulfonamides in foods. Recovery studies yielded good results for all food samples because there were no interferences from the matrices.
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  • 44
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    Chromatographia 40 (1995), S. 411-416 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Particle beam LC/MS interface ; Capillary columns for LC ; Mycotoxins ; Peanut meal
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A new method for the analysis of aflatoxins in food extracts, based on liquid chromatography/mass spectrometry interfacing, is presented. The chromatographic separation was performed with a reversed phase packed capillary column coupled with a modified particle beam interface capable of handling microliter per minute flow rates. This system allows higher overall sensitivity and easier operation procedures. The method has proved to be particularly suitable for the analysis of the toxins in very complex matrices. The specificity of electron impact ionization allowed positive identification of the aflatoxins with an excellent response linearity for accurate quantitation.
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  • 45
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Thin-layer chromatography ; Reversed phase ; 2-benzoylbenzoic acids ; Anionic and cationic counter ions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The 2-benzoylbenzoic acid series was investigated by reversed-phase, high-performance, thin-layer and column chromatography using various alkylammonium salts and di(2-ethylhexyl)orthophosphoric acid as polar associating reagents. The effects of the individual substituents on retention were quantified by Δlog k′ and ΔRM values. The compounds investigated differing in molecular structure (hydrophilic and hydrophobic substituents) commonly occurring groups in drugs and biologically active substances provide information on molecular interaction in these ion-pair systems. The combined effects on retention of organic modifier and ion-pair reagent concentration were investigated.
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  • 46
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Two-dimensional separation ; D- and L-amino acids ; Enantioseperation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A two-dimensional, column liquid chromatographic system is used for the determination of the D- and L-enantiomers of amino acids in biological samples. Separation of the amino acids is first on ion-exchange column by gradient elution with a sodium citratesodium chloride buffer. Enantioseparation is by subsequent injection of 3 μl heart-cuts of the individual amino acids onto a second column with a chiral crown ether stationary phase. Finally, fluorescence detection is after post-column labelling of the amino acids using ano-phthalaldehyde-2-mercaptoethanol reagent solution. The high separation power and selectivity of the system allow processing of complex samples with hardly any additional treatment and the determination of small quantities of D-amino acids in the presence of excess L-form. Applicability of the system is illustrated by the determination of D- and L-aspartate, serine, glutamate and alanine in various complex biological samples, such as protein hydrolysates, urine and biotechnological and food samples. Data are given on detectability, repeatability and linearity.
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  • 47
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ion pair LC ; Sweeteners, preservatives and antioxidants ; Food additive analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A paired-ion, reversed-phase high-performance liquid chromatographic method was developed for the simultaneous determination of sweeteners (dulcin, saccharin-Na and acesulfame-K), preservatives (sodium dehydroacetate, sorbic acid, salicyclic acid, benzoic acid, succinic acid, methyl-para-hydroxybenzoic acid, ethylpara-hydroxybenzoic acid, n-propyl-para-hydroxybenzoic acid, isopropyl-para-hydroxybenzoic acid, n-butyl-para-hydroxybenzoic acid, and isobutyl-para-hydroxybenzoic acid), and antioxidants (3-tertiary-butyl-4-hydroxyanisole and tertiary-butyl-hydroquinone). A mobile phase of acetonitrile-50 mM aqueous α-hydroxy-isobutyric acid solution (pH 4.5) (2.2 ∶ 3.4 or 2.4 ∶ 3.6, v/v) containing 2.5 mM hexadecyltrimethylammonium bromide and a C18 column with a flow rate at 1.0 mL/min and detection at 233 nm were used. This method was found to be very reproducible with detection limits ranged from 0.15 to 3.00 μg. The retention factor (k) of each additive could be affected by concentrations of hexadecyltrimethylammonium bromide and α-hydroxyisobutyric acid, and pH and ratio of mobile phase. The presence of additives in some food samples was determined.
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  • 48
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Limit of detection ; Precision ; Uncertainty prediction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The precision of integration over noisy instrumental output for quantitative analysis is studied. A probability theory is developed to predict the relative standard deviation (RSD) of integration results over an integration domain from one-point integation (peak height measurement) to entire area integration in HPLC. Common integration modes of horizontal zero line and oblique zero line are taken into account, but no peak overlap is assumed. The question of the analytical superiority of peak height measurement or integration for quantitation is answered. In the HPLC apparatus used, the minimum RSD of measurements is found in the integration domain of ca. ±0.5 σ for analytes [peaks are approximated by the Gaussian signal of width, σ (standard deviation)]. The RSD of integration measurements is also shown to depend on the stochastic properties of back-ground noise (uncorrelated noise and correlated 1/f type noise). The theoretical conclusion is verified by Monte Carlo simulation and HPLC experiments for some aromatic compounds.
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  • 49
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ion-pair and GPC separations ; Amoxicillin oligomers ; β-Lactam ring polymers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Polymeric substances formed from concentrated sodium amoxicillin in an aqueous solution have been separated using two high-performance liquid chromatographic methods. We used a C18 reversed-phase column with tetrabutylammonium chloride as an ion-pairing agent with an acetonitrile gradient and a TSKgel G2500PWxl column with water as the solvent for gel permeation chromatography. The separated materials, ranging in size from the monomer to the tetramer, have been characterized by functional-group chemical analysis, while the identification of the piperazine-2,5-dione was done using a pure standard. A greater number of peaks which were also better defined were obtained using the ion-pair reversed-phase method, and open and closed beta-lactam ring polymer forms could be distinguished. Using the gel permeation method, only a few monomer, piperazine-2,5-dione, dimer, trimer and combined amoxicillin trimer and tetramer peaks were obtained with water, although those obtained were quite well defined. The data on the time-course of formation of the oligomers and the amoxicillin degradation product were virtually identical by both methods.
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  • 50
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; On-column fluorimetric detection ; Gradient elution ; Dansyl amino acids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Fluorimetric detection in the presence of a stationary phase has been applied to gradient elution of dansyl amino acids in liquid chromatography. A 1.5 mm ID quartz tube packed with the same materials as the separation column was employed for the flow cell. Conventional-size columns were employed. The peak height of analytes increased with increasing retention owing to focusing and environmental effects of the stationary phase, leading to improvements in sensitivity, which was pronounced for analytes eluting late. The lower the gradient, the larger the improvement in sensitivity achieved. Detection limits were improved by a factor of up to 5.1 by fluorimetric detection using the packed flow cell, compared with those achieved using a common empty flow cell.
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  • 51
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; LC-MS ; Cinchona alkaloids ; New column packing materials
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A major problem in the HPLC analysis of alkaloids is the poor peak shape and consequently low resolution, due to the interactions of the basic alkaloids with the residual acidic silanol groups of most reversed phase materials. The performance of new packing materials specially designed for the separation of basic compounds has been studied using mobile phases without the special additives commonly applied in the analysis of alkaloids. Strongly basic Cinchona alkaloids were used as test compounds. Retention characteristics and selectivities of each material were studied, after mobile phase optimisation for the column. The influence of the major factors (nature and content of the organic modifier, pH value, salt concentration) affecting resolution was studied. The mobile phases were chosen so that they could be used in thermospray LC-MS. The addition of salts to the mobile phase improves separation but in general the modification of the mobile phase gave little change in selectivity. The performance of silicabased C18 material proved superior to the polymer materials tested.
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  • 52
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    Protoplasma 184 (1995), S. 86-103 
    ISSN: 1615-6102
    Keywords: Neutrophil ; Leukocyte ; NADPH ; Oxidase ; Microbicidal ; pH
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Phagocytic cells of the immune system contain an oxidase that is important for the killing and digestion of engulfed microbes. This is an electron transport chain that transfers electrons from NADPH in the cytosol to oxygen to form superoxide and hydrogen peroxide in the phagocytic vacuole. Absence or abnormality of this oxidase results in the syndrome of CGD, characterised by a profound predisposition to infection. The electron transport chain consists of a flavocytochrome b located in the plasma membrane and membrane of the specific granules. It is composed of a and b-subunits, with apparent molecular masses of 23 kDa and 76–92 kDa, respectively. The b-subunit is a member of the FNR family of reductases with FAD and NADPH binding sites. Based upon the crystal structure of FNR we have constructed a model of the more hydrophilic C terminal half of this b-subunit, which acts as a guide to the organisation of the molecule, and provides a template on which to map mutations in CGD. The location of the heme is uncertain. Electron transport is dependent upon an activation complex of cytosolic proteins including p40 phox , p47 phox , and p67 phox , and the small GTP binding protein, p21 rac . This oxidase system is important for the killing and digestion of bacteria and fungi. This might be accomplished in a number of ways. The oxidase produces superoxide and hydrogen which might be toxic themselves. The hydrogen peroxide can act as substrate for myeloperoxidase which can oxidise chloride and iodide to chlorine and iodine and their hypohalous acids. The proteins contained within the cytoplasmic granules are also very important in the killing process. These are neutral proteinases that require a neutral or slightly alkaline pH for optimal activity. The oxidase transports electrons, unaccompanied by protons, across the wall of the phagocytic vacuole, resulting in an elevation of the vacuolar pH, thereby optimising conditions for killing and digestion of engulfed organisms by these neutral proteinases.
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  • 53
    ISSN: 1615-6102
    Keywords: Acridine orange ; Carrot ; Embryogenesis ; Neutral red ; pH
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The accumulation of neutral red and acridine orange, to indicate differences in vacuolar pH, was studied during embryogenesis of carrot. Neutral red accumulated barely in proembryogenic masses, but was present conspicuously in globular-shaped somatic embryos. From the late globular to the torpedo-shaped stage, it was mainly found in the root side of the somatic embryo. Here, neutral red was predominantly present in large dark-red to purple stained vesicles. In the cotyledons neutral red was found in small orange vesicles. In zygotic embryos of carrot, the dye was uniformly distributed with no specific localization in organelles. During germination, however, neutral red accumulated mainly in regions in the root side and the hypocotyl of the germling. Acridine orange was dispersed erratically in proembryogenic masses with a great variety in intensity. It was quite obviously present in early stages of somatic embryogenesis and restricted to the root side in late globular to torpedo-shaped embryos. Confocal images revealed the vacuolar presence of the fluorescence and the predominant presence in the protoderm. During germination of zygotic embryos the signal changed from uniform to localized, with sharp borders between fluorescent and non-fluorescent regions. Two to three days after the beginning of germination, acridine orange accumulated preferentially in the root tip of the germling. Differences between somatic and zygotic embryos and similarities between somatic embryogenesis and zygotic embryo germination are discussed.
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  • 54
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    Aquaculture international 3 (1995), S. 123-133 
    ISSN: 1573-143X
    Keywords: Ammonium ; Coastal ponds ; Hydrogen sulphide ; pH ; Redox potential ; Sediment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Aquaculture ponds receive inputs of organic matter such as uneaten feed, fertilizer and faeces. Analytical methods for four parameters are described to assess the sediment quality. These parameters are redox potential, pH, the hydrogen sulphide activity potential (pH2S) and soluble ammonium nitrogen. The techniques are simple, rapid, and are practical on site. Ranges encountered are given and ‘safe’ ranges suggested.
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  • 55
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    Journal of plant research 108 (1995), S. 249-256 
    ISSN: 1618-0860
    Keywords: Characeae ; H+ pump ; pH ; Pi ; Plasma membrane ; Tonoplast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The relationship between the physiological characteristics and changes in the activities of H+ pumps of the plasma membrane and tonoplast of characean cells is discussed. The large size of the characean internodal cells allows us to perform various experimental operations. The intracellular perfusion technique developed by Tazawaet al. (1976) is a powerful tool for analyzing the characteristics and control mechanisms of the H+ pumps (Tazawa and Shimmen 1987, Tazawaet al. 1987, Shimmenet al. 1994) Respiration-dependent changes in the activity of the plasma membrane H+ pump are explained by changes in the supply of energy substrate. Photosynthesis-dependent changes in activities of both the plasma membrane and the tonoplast H+ pumps are explained in terms of changes in the level of inorganic phosphate in the cytoplasm. Cytoplasmic and vacuolar pHs are suggested to be controlling factors forin vivo activities of the H+ pumps. Furthermore, the membrane potential and various ions are considered to bein vivo factors that regulate the activities of the H+ pumps.
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  • 56
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    Plant and soil 171 (1995), S. 175-178 
    ISSN: 1573-5036
    Keywords: aluminium ; mycorrhizae ; pH ; root growth ; X-ray microanalysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Norway spruce (Picea abies) was exposed to nutrient solutions containing a range of aluminium (Al) concentrations at several pH levels (3.2, 4 and 5). Root growth was reduced by 100 µM and 400 µM Al at pH 4 and 5, but at pH 3.2 only by 400 µM Al. The Al content of the roots increased with increasing pH. The Al content of the roots was higher at the root tips than at the older root parts at all pH values. Using X-ray microanalysis it could be shown that higher levels of Al at increased pH were mainly due to increased Al contents in root cortex cell walls. In seedlings, mycorrhizal with Pisolithus tinctorius or Lactarius rufus, the Al concentration of cortex cell walls was higher when nitrate (NO3) rather than ammonium (NH4) was the nitrogen (N) source.
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  • 57
    ISSN: 1573-5036
    Keywords: aluminium ; organic acids ; pH ; stubble burnt ; stubble retained
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Soil solution and solid phase analyses were compared for stubble standing and stubble burnt systems on an acid, acidified and limed Alfisol in north-eastern Victoria. Exchangeable (1 M KCl) aluminium (Al), soil solution total and monomeric Al, pH (water) and soil solution pH were measured to identify any differences in soil acidification between the stubble burnt and the stubble standing treatments. All measures of Al increased with soil depth down to 10 cm in all stubble treatments and decreased in the 10–20 cm soil depth with the exception of lime treatments. Concentrations of low molecular weight organic acids in the soil solution were determined by high pressure liquid chromatography (HPLC). Aromatic and aliphatic acids were distributed throughout the soil depth (0–20 cm) for the stubble standing treatments but were confined to the 0–5 cm soil depth for the stubble burnt treatment. In this short term field trial (2 years for stubble management), the data suggest that the conventional practice of stubble burning was more acidifying than the soil conservation practice of stubble standing, over the 0–20 cm soil depth for the lime (6 t ha-1) treatments.
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  • 58
    ISSN: 1573-5036
    Keywords: distribution ; germination ; H+ ; pH ; soil solution ; toxicity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Deschampsia flexuosa (L.) Trin. is an abundant grass species in the ground flora of acidic beech forests in southern Sweden. Generally, the species is restricted to a limited soil pH range (pH 4–5). The main objective was to study the influence of different soil acidities on germination, initial root development and on the growth of the species. The experiments were carried out under controlled conditions and designed to simulate the physico-chemical conditions present in the field. By using forest soils within the pH range 4.0 to 8.3 and artificial variation in pH (3.2 to 7.6) of soil-water extracts, it was possible to evaluate the influence of soil reaction and the H+ per se. In all experiments seeds have been used. Germination was significantly delayed in the very acid soil (pH 4.0) in comparison to the germination in soils within the pH range (4.4 to 6.4). Soil substances, other than the H+, might be responsible for this delay in germination, whereas development of the radicle was markedly affected by increasing H+ concentrations. Especially the development of root hairs was sensitive to H+ and was significantly reduced at a pH〈-3.8. By increasing soil acidity the injury symptoms, including curling and discolouring, became more intense and at the highest acidity (pH 3.2) the radicles appeared brown, stunted and the root hairs were lacking. Most favourable growth was obtained at pH 4.4 and 5.0. Soil pH levels above and below this range limited both shoot and root growth. The results showed very good correspondence with observations made in Beech forest soils in southern Sweden, where the species was growing in soils within the pH range 3.9 to 5.1 with a peak growth at pH 4.3. This study shows that in soils at pH≤3.8, the poor development of the radicle may be crucial in the establishment ofDeschampsia flexuosa. Root hair development was more sensitive to soil acidity than radicle elongation. Germination was delayed in very acid Beech forest soils but other factors than the H-ion per se may be responsible for this delay.
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  • 59
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    Plant and soil 168-169 (1995), S. 55-65 
    ISSN: 1573-5036
    Keywords: acidity ; autotro[hic nitrifiers ; N mineralization ; nitrification ; pH ; soil horizon
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Soil samples were collected from the litter, humus and five mineral soil layers to a depth of 50 cm in acid, but highly productive, Norway spruce forests in S Sweden and E Denmark for determination of net N mineralization and potential nitrification. The samples were sieved while still fresh and incubated at a constant temperature (15°C) and soil moisture for 74–117 days with periodic subsamplings. Net N mineralization rates, expressed per g organic matter or per g of total N, decreased with increasing depth. Net N mineralization in the ten soils studied ranged from 35 to 105 kg N ha- yr-1, of which the organic horizons contributed 32–74%. Nitrate formation patterns were variable. Almost no nitrification could be detected at pH (H2O) values lower than 4.0. Nitrate was formed in humus layers with pH values of 4.0–4.5, but the nitrification was never complete. By contrast, the nitrification was almost complete at a depth of 10–50 cm, where the pH (H2O) was 4.1–4.5. Addition of CaCO3 stimulated nitrification in the humus layer, indicating the presence of acid-sensitive nitrifiers, while nitrification in the mineral soil was sometimes sthnulated and sometimes inhibited by the addition of CaCO3. Tests with the acetylene block method showed that nitrification, when it occurred, was autotrophic in the humus layer but less easy to characterize in the mineral soil. Thus, further studies are needed to characterize the nitrifier organisms and identify the factors regulating the activity of these organisms. Actual nitrification rates in the field could not be determined, but our findings showed that the B horizon in acid forest soils has a great potential for nitrification. Because N deposition does not appear to decrease in Scandinavian forests, we conclude that an ammonium surplus in the forest floor followed by a downward transport of ammonium to the nitrifying subsoil can increase the risk of nitrate leaching in the future.
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  • 60
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    Plant and soil 168-169 (1995), S. 103-110 
    ISSN: 1573-5036
    Keywords: afforestation ; carbon ; cation exchange capacity ; Norway spruce ; pH ; soil changes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The aim of this study was to test the hypothesis that afforestation changes the content and distribution of soil organic carbon, nutrients and pH in the A-horizon of land previously used in agriculture, and that such soil changes depend on stand development. The investigation was evaluated as a completely randomised design with three treatments representing different age classes of trees: 20 years (Y20), 40 years (Y40) and 55 years (Y55). Eighteen trial plots, six per treatment, were established in plantations of Picea abies (L.) Karst. on soils of similar texture and mineralogy. Tree volume was 220 m3 ha-1 in Y20, 400 in Y40 and 440 m3 ha-1 in Y55. Concentrations of carbon (C) and nitrogen (N) were significantly higher in the uppermost part of the soil in the older stands Y40 and Y55 than in Y20. The total amount of organic C in the litter layer plus the top 15 cm of the soil differed between age classes, with Y40 and Y55 having the largest amounts. A reference layer (15–20 cm) was used in calculating the amount of soil C that had accumulated in the horizon since afforestation, being about 10 tonnes ha-1 of C in Y20 and 19 tonnes ha-1 in Y40 and Y55. Cation exchange capacity (CEC) and base saturation (BS) was higher in the older stands. Carbon contents and CEC were strongly correlated. In Y40 and Y55, pH was significantly lower than in Y20 in the lower part of the soil horizon. There was a general decrease with depth of C, N, CEC, K+ and Mg2+ in the soil horizon. BS, Ca2+, Na+ and pH showed a somewhat different pattern of distribution, with deceasing values in the upper part of the soil horizon and increasing values in the lower part of the soil horizon.
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  • 61
    ISSN: 1573-5036
    Keywords: Ca:Al ratio ; exchangeable Ca and Al ; forest plant distribution ; pH ; soil solution Ca and Al
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The distributions of vascular plants in south Swedish deciduous forests were related to exchangeable (exc) and soil solution concentrations of H+ (pH), Ca, Al and the Ca:Al ratios within these fractions. Topsoils (0–5 cm) of 172 sites with a pHKCl of 3.2–3.9 (corresponding to 3.7–4.4 in soil solution) were used. In the soil solution both total Alt and quickly reacting Alr were determined. Exchangeable concentrations were generally well related to plant distributions, the highest correlation coefficients usually being given by pHKCl〉Caexc〉Alexc.〉(Ca:Al)exc. The (Ca:Al)exc ratio was clearly inferior. Out of the soil solution variables studied, Ca concentration, followed by pH, was best correlated with plant distributions, Alt, Alr, and the Ca:Al ratios having similar and lower coefficients. It is concluded that the use of Ca:Al ratios as a general measure of Al toxicity in controlling plant distributions is rather problematic. It seems difficult to apply evidence for Ca-Al interactions from solution culture experiments to field conditions when measured as exchangeable or soil solution concentrations of the soil.
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  • 62
    ISSN: 1573-5036
    Keywords: aluminium ; episodes ; flowing solution culture ; pH ; reversibility, toxicity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Herbs and grasses may episodically be exposed to elevated concentrations of potentially toxic elements as H+ and Al3+ due to considerable temporal variability in the chemical composition of soil solutions in the uppermost layer of the soil profile. Greenhouse experiments were performed to test the effect of episodes of different length (1 and 2 weeks) and concentrations of H+ and Al3+ (pH 3.8, 4.0, 4.2 and 4.5; 0, 20 and 70 μM Al) on root and shoot growth, designed to elucidiate the reversibility of growth inhibition. Three forest floor species were studied-Galium odoratum and Lamium galeobdolon, having similar pH distribution ranges in the field and Poa nemoralis which also occurs at slightly lower pH. The plants were grown for 5 weeks (episode and recovering time) in a synthetic soil solution in a flowing solution system without recirculation. The species reacted in three different ways. Galium odoratum was the most sensitive species and seemed to be irreversibly damaged (ceased growth) by 2 week episodes of pH≤4.0 and Al≥20 μM at pH≤4.2. Lamium galeobdolon was about equally sensitive during the episodes but it had a much greater ability to recover. Poa nemoralis was rather insensitive to the episode treatments tested. It is concluded that episodes of elevated H+ and Al3+ concentrations may be decisive for plant performance. It is therefore important to consider the extreme environmental conditions which plants may be exposed to in the field, in addition to long-term averages of e.g. soil solution concentration of potentially toxic elements, when studying species distribution and performance in relation to soil chemical properties.
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  • 63
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    Plant cell, tissue and organ culture 43 (1995), S. 123-126 
    ISSN: 1573-5044
    Keywords: bioreactor ; dissolved oxygen ; haploid cell suspension ; pH ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Maize (Zea mays L.) haploid cells were cultivated in a 1500 ml aerated and stirred batch bioreactor using modified BM medium. Cell growth was highly affected by pH and dissolved oxygen, and we observed two fairly distinct growth phases. During the first two days after inoculation at pH 5.8, oxygen consumption was high and the cells lowered the pH to a value around 4.3. After this period the pH stabilized at 4.5 and the dissolved oxygen reached a steady level. Decreasing dissolved oxygen concentration leads to lower growth rate and to higher pH. Both events mean stress conditions for the cell culture and probably result in increased genetic variability, and the loss of regeneration capacity. The stress condition during the adaptation phase can be eliminated by decreasing the pH of the medium to 4.7 before inoculation and by keeping dissolved oxygen above 40%. These conditions provide prolonged exponential growth dynamics and the cell suspensions could be the basis of large scale cultures also.
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  • 64
    ISSN: 1435-1536
    Keywords: Adsorption ; codeine ; surface modified ; silica ; alkyl group ; electrolyte ; pH
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: Abstract The adsorption of codeine from aqueous solution onto colloidal silica and silica surface-modified with chemiadsorbed octadecyl dimethyl silane (ODDMS) or dimethyl silane (DMS) groups was studied in the presence of neutral electrolytes at different pH values. From codeine-hydrochloride solutions codeine cations are strongly bound to negatively charged silica surfaces. Inorganic salts (NaCl, NaNO3) reduce the adsorption of the organic cation. On silica modified by ODDMS (10% of surface silanol groups are occupied), codeine cations are adsorbed to a higher extent at pH 6, while at pH 8 the adsorbed amounts are lower than on the bare silica surface. Neutral electrolytes reduce codeine adsorption on the ODDMS modified silica. On the hydrophobic silica, completely covered by DMS groups, codeine adsorption is considerably lower than on the bare silica, but neutral salts increase the adsorption. The adsorption of codeine is compared with the adsorption of aggregating surfactant ions. Common and different features of their interactions with silica surfaces are outlined.
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  • 65
    ISSN: 1573-5079
    Keywords: cytochrome b 559 ; electron transport ; pH ; pheophytin ; photosynthesis ; Photosystem II ; reaction center ; redox potential
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A series of experiments have been conducted with isolated reaction centers of photosystem two (PS II) with the aim to elucidate the functional role of cytochrome (Cyt b 559). At pH 6.5 it was found that Cyt b 559 was reversibly photoreduced by red actinic light when Mn2+ was present as an electron donor while at pH 8.5 a photo-oxidation was observed under the same lighting conditions, which was dark reversible in the presence of hydroquinone. These pH dependent light induced changes were measured under anaerobic conditions and correlated with changes in the relative levels of high (HP) and low (LP) potential forms of the cytochrome. At pH 6.5 the cytochrome was mainly in its LP form while at pH 8.5 a significant proportion was converted to the HP form as detected by dark titrations with hydroquinone. This pH dependent difference in the levels of HP and LP Cyt b 559 was also detected when bright white light was used to monitor the level of the LP form using a novel reaction involving direct electron donation from the flavin of glucose oxidase (present in the medium and used together with glucose and catalase as an oxygen trap). The results suggest that PS II directly oxidises and reduces the HP and LP forms, respectively and that the extent of these photo-reactions is dependent on the relative levels of the two forms, which are in turn governed by the pH. This conclusion is interpreted in terms of the model presented previously (Barber J and De Las Rivas J (1993) Proc Natl Acad Sci USA 90: 10942–10946) whereby the pH induced effect is considered as a possible mechanism by which interconversion of LP and HP forms of Cyt b 559 is achieved. In agreement with this was the finding that as the extent of photo-oxidisable HPCyt b 559 increases, with increasing pH, the rate of irreversible photo-oxidation of β-carotene decreases, a result expected if the HP form protects against donor side photoinhibition.
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  • 66
    ISSN: 1573-2932
    Keywords: Precipitation chemistry ; sulfate ; nitrate ; ammonium ; calcium ; pH ; Japan
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Precipitation chemistry in Japan was discussed on a wet-only sample database obtained in a nationwide survey from April 1989 to March 1993. Wet-only samples were collected at 29 stations over Japan on a biweekly basis. Commonly determined chemical parameters were measured in laboratories. The volume-weighted annual mean pH at each site ranged from 4.50 to 5.83 with a mean of 4.76. Concentration ranges and means (parenthesized) on an equivalent basis for major ions were as follows: nss-SO4 2−; 5.2–58.9 (38.6), NO3 −; 1.8–25.0 (14.1), NH4 +; 0.55–29.8 (18.3), nss-Ca2+; 2.0–34.5(14.2), Na+; 6.4–275.3 (49.1), Cl−; 13.7–322.4 (63.5) β eq L−1. Acid-base relationships for Phase-II records were quantitatively discussed in terms of three measures: pH, fractional acidity, and our proposed pAi.
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  • 67
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    Water, air & soil pollution 85 (1995), S. 1985-1990 
    ISSN: 1573-2932
    Keywords: cloud-water ; sulphur dioxide ; oxidation ; catalysis ; trace metals ; iron ; acidity ; pH
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Aerobic oxidation of S(IV) was investigated in cloud-water samples collected at Great Dun Fell,U.K., as part of a wider project into cloud-water chemistry. The rate was found to be first-order in S(IV) concentration, and the reaction rate constant. kS(IV), was found to vary from 10−5 to 10−3 s−1. The rate constant was highly correlated with H+ concentration (pH 3.5 to 6.5) and Fe concentration (〈0.02 to 3×10−6 mol dm−3). The aerobic oxidation of S(IV) does not contribute significantly to SO2 oxidation in clouds at Great Dun Fell, however, the reaction may be of consequence in clouds and fogs at polluted or urban sites with elevated trace metal concentrations. Also, this reaction may be responsible for the oxidation of S(IV) in cloud-water samples during storage.
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  • 68
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    Ecotoxicology 4 (1995), S. 299-306 
    ISSN: 1573-3017
    Keywords: pH ; aluminium ; Triturus ; growth ; feeding behaviour
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: The distribution of smooth and palmate newts (Triturus vulgaris and Triturus helveticus) may be related to water quality. This study compared the larval growth and feeding behaviour of both species under sublethal levels of aluminium and low pH. The growth of both species was inhibited to a similar degree under acidic conditions with aluminium toxicity dependent upon both low pH and developmental stage. The two species were differentially affected by low pH in terms of feeding behaviour. T. vulgaris larvae took significantly longer to snap at food under low pH than the apparently acid-tolerant T. helveticus. Aluminium had no significant effect on the feeding behaviour of either species.
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  • 69
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Diode-array detection ; Retinol acetate ; Pharmaceutical preparations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A very simple high-performance liquid chromatographic method for determination of Vitamin A in pharmaceutical preparations without the need for saponification was developed. A reversed-phase (Nova-Pack C18, 4 μm) column was used with a mobile phase of acetonitrile-tetrahydrofuran-water (55∶37∶8) and a flowrate of 1.5 ml/min. Sample treatment only consisted of the extraction of retinol acetate content from capsules or tablets with methanol. Total extraction was achieved by shaking vigorously with the aid of magnetic stirring for three hours at room temperature. No change of solvent is necessary to introduce the sample in the chromatographic system. This method is suitable for routine quantification of Vitamin A.
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  • 70
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    Chromatographia 40 (1995), S. 193-196 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Chemometric methodology ; Elution order ; Simplex optimization ; Benzodiazepines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Using a rapid chemometric methodology to determine the separation factor, α, at different temperatures, Gibbs Helmholtz parameters (Δ (ΔH), Δ (ΔS), Δ (ΔG)) of two adjacent benodiazepines on a chromatogram were obtained from ln α versus T−1 plots. A temperature dependent reversal of the elution order was studied and the mobile phase composition and column temperature were optimized to obtain the best separation. A flow rate of 0.80 ml min−1 with 52.6% methanol in the methanol-water mixture and a column temperature of 48°C gave the most efficient separation of ten benzodiazepines.
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  • 71
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    Chromatographia 40 (1995), S. 237-246 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Mixed mode stationary phases ; Indirect photometric and fluorometric detection ; Conductimetric detection ; Alkyl-sulphates, and sulphonat-phosphates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Separation of three classes of anionic surfactants (alkyl sulfates, alkanesulfonates, and alkyl phosphates) are achieved on a mixed-mode reversed phase (RP) phenyl/-anion exchange column using a naphthalenetrisulfonate (NTS)-acetonitrile (ACN) mobile phase via indirect photometric, indirect fluorometric, direct or indirect conductivity detection. Mixtures of C5−C18 sulfates, C6−C18 sulfonates, and C1−C4 phosphates (mono- and di-ester) can be separated in less than 20, 24, and 20 min respectively. Although hydrophobic effects are more pronounced in mixed-mode chromatography, equivalent-per-equivalent exchange of analysis and eluent ion is still required for sensitive indirect photometric, fluorometric, or conductivity detection. The detection limits of alkyl sulfates and alkanesulfonates are in the range of 3–15 pmoles which are at least an order of magnitude better than suppressed conductivity detection. The determination of surfactants in a variety of real samples is also presented.
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  • 72
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    Chromatographia 40 (1995), S. 287-295 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Capillary zone electrophoresis ; Enantioseparation ; Carnitine ; Derivatization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Carnitine is an essential component in tissues of animals, higher plants and many microorganisms. Whereas the L-carnitine enantiomer plays an important role in the metabolism of long chain fatty acids, D-carnitine has a considerable toxic influence on biochemical processes. The analytical separation of D-and L-carnitine depends upon derivatization with UV-or fluorescently active substances, e.g. FMOC and (+)/(−)-FLEC. The separation of diastereomeric (+)- and (−)-FLEC carnitine esters was performed successfully with capillary zone electrophoresis (CZE) and HPLC, after optimization of the derivatization process and of the composition and pH of the buffer, using UV- and fluorescence detection. With HPLC separation a detection limit of the carnitine esters of 5 μmol/l when using fluorescence detection was achieved. With both separation systems baseline resolution and short analysis times could be obtained. The enantiomeric FMOC derivatives could be separated using the electrophoretic system and acidic buffers with high concentrations of an osmotic flow modifier together with γ-cyclodextrine as chiral selector. The applicability of the optimized separation conditions are demonstrated in the analysis of agar culture medium inoculated withPseudomonas putida and of pharmaceutical formulations. In all samples very low amounts of D- or L-carnitine could be determined in the presence of the other enantiomeric form. Problems caused by the impurity of the carnitine standards or the derivatization agent (+)/(−)-FLEC are discussed.
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  • 73
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Fullerenes ; Retention vs NMR spectra ; C18 phases ; Low temperature separations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The temperature effect on the retention of fullerenes in the range 80°C to −70°C in liquid chromatography (LC) has been examined using octadecylsilica stationary phases (ODS). It has been found that the maximum retention temperature lies around −10°C with a highly carbon loaded ODS phase. Solid state CPMAS NMR measurements on the stationary phases indicated that the relaxation time at the 30 ppm methylene signal changes with the temperature and has a minimum relaxation time at the temperature which closely matches the maximum retention temperature observed in chromatography. The interpretation of both NMR spectroscopic and LC chromatographic data are discussed.
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  • 74
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    Chromatographia 40 (1995), S. 601-606 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Amino acid analysis ; Chirality ; Diastereoisomers ; Dipeptides
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Using a robotic autosampler, free DL-amino acids were derivatized with highly reactive, urethane-protected, L-α-amino acidN-carboxyanhydrides (NACs) with structures: Boc-Phe-NCA, Boc-Asn(Trt)-NCA, Fmoc-Lys(Boc)-NCA, Fmoc-Met-NCA, Fmoc-Ala-NCA, Z-Ala-NCA, Z-Val-NCA and Z-Leu-NCA (Boc=tert-butyloxycarbonyl, Trt=trityl, Fmoc=9-(fluor-enylmethyl)oxycarbonyl, Z=benzyloxycarbonyl). Using sodium borate buffer and acetonitrile solvent, derivatization was complete in 3.5 min at room temperature. By selection of an appropriate reagent, the resulting diastereomeric,N-protected dipeptides were separated on an octylsilica stationary phase using mixtures of sodium acetate buffer and acetonitrile as eluents.
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  • 75
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Dihydroxyacetone ; 2,4-dinitrophenylhydrazine ; Tanning creams
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A rapid and simple HPLC method for the identification and determination of dihydroxyacetone, a tanning agent widely used in a variety of preparations, is presented. The method consists of the analysis of the formulated products after a derivatization reaction which enhances the response and the detection limit of the chromatographic analysis. The characterization and the elucidation of the spacial geometry of the synthesized derivative are described.
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  • 76
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    Chromatographia 41 (1995), S. 431-434 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Thin-layer chromatography ; Patent Blue V in cheese
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The HPLC method developed for the analysis of the dye Patent Blue V in extracts from cheese is sufficiently sensitive to detect and measure concentrations above 0.1 ppm with a standard deviation of 3%. The extraction procedure described gives a recovery from cheese of about 80%. The method has been applied to commercial samples of cheese and a concentration of the dye of about 0.12 ppm was measured in one case.
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  • 77
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Micellar liquid chromatography ; Neural networks ; Retention modelling
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Multi-layer feed-forward neural networks trained with an error back-propagation algorithm have been used to model retention behaviour of liquid chromatography as a function of the composition of the mobile phases. Conventional hydro-organic and micellar mobile phases were considered. Accurate retention modelling and prediction have been achieved using mobile phases defined by two, three and four parameters. With micellar mobile phases, the parameters involved included the concentrations of surfactant and organic modifier, pH and temperature. It is shown that neural networks provide a competitive tool to model varied inherent nonlinear relationships of retention behaviour with respect to the mobile phase parameters. The soft models defined by the weights of the networks are capable of accommodating all types of linear and nonlinear relationships, neural networks being specially useful when the relationships between retention behaviour and the mobile phase parameters are unknown. However, to train neural networks more experimental points than with hard-modelling methods are required, hence the use of the networks is recommended only for those cases where adequate theoretical or empirical models do not exist.
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  • 78
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Diltiazem hydrochloride stereoisomers ; Chiralcel OF ; Chiral inversion ; Epimerization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The simultaneous stereospecific assay of four stereoisomers of diltiazem hydrochloride in bulk drug and aqueous solution was developed using HPLC on a Chiralcel OF column. The four isomers were quantitated with good precision by the internal standard method. The chiral inversion of (+)-cis-diltiazem hydrochloride in vitro, stability of its (2S, 3S) configuration in the solid and aqueous states was examined by HPLC. Chiral inversion of (+)-cis-diltiazem hydrochloride was not observed in the solid state, and its (2S, 3S) configuration was stable to heat, humidity and light. Chiral inversion of (+)-cis-diltiazem hydrochloride (2S, 3S) was observed in aqueous solution under UV, but not in aqueous solution stored at 80°C for 5h nor under visible light for 10 h. The (+)-cis-diltiazem hydrochloride (2S, 3S) epimerized to (+)-trans-diltiazem hydrochloride (2R, 3S) with a half-life of 5h in aqueous solution under UV but the reverse chiral inversion of (+)-trans-diltiazem hydrochloride (2R, 3S) to (+)-cis-diltiazem hydrochloride (2S, 3S) was not observed.
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  • 79
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    Chromatographia 41 (1995), S. 488-491 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Acetonitrile disposal ; Acetonitrile hydrolysis ; Degradation kinetics ; Waste solvent management
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Acetonitrile is one of the most frequently used solvents in column liquid chromatography. Because of its toxicity, safe disposal of the waste solvent is essential. Combustion in a hazardous waste plant, if available, is recommended. Otherwise the waste can be degraded in the laboratory to acetic acid and ammonia by treatment with sodium hydroxide. The addition of hydrogen peroxide is not necessary. The waste needs to be diluted to 10% acetonitrile in water in order to prevent the formation of a two-phase system on addition of concentrated sodium hydroxide. This reagent must be added in excess; 2.5 mols of sodium hydroxide per mol of acetonitrile are recommended. It was found that the kinetics are of second order, with an activation energy of 89 kJ mol−1 and a frequency factor of 9×109. Therefore even at room temperature degradation is possible, although a hydrolysis time of approximately 15 days is needed under these conditions. At 80°C, where refluxing is not necessary, hydrolysis is complete after, e.g., 2 h, depending on the amount of sodium hydroxide added.
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  • 80
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ion-pair LC ; 5-Sulphoquinoline-8-ol ; Metal complexes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The reversed-phase ion-pair liquid chromatographic behaviour of several metal complexes with 5-sulphoquinoline-8-ol (HQS) on an octadecyl-modified silica column was investigated by using a fluorescence detector. The aluminum complex gave two peaks on the chromatograms when eluents containing phosphate buffer were used. The effects of the pH of the eluent, HQS concentration, the nature of anions in the eluent, and the column temperature on the retention behaviour of the aluminum complex are discussed on the basis of the equilibria and the kinetics of the complexation of the aluminum ion with HQS and anions in the eluent. The equilibrium constant for the interconversion reaction between the two Al(III)-HQS species which correspond to the two peaks observed and the second stepwise formation constant for the Zn(II)-HQS complex were evaluated from the HPLC data obtained.
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  • 81
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Fluorescamine derivatives ; Aniline and aniline derivatives ; Environmental waters
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Fluorescamine (fluram) has been used as a fluorogenic compound for pre-column derivatization of aniline and some derivatives. Anilines were derivatized with fluram in citrate buffer media (pH 5.5) to form pyrrolinones. The highly fluorescence pyrrolinones were isolated and pre-concentrated by solid phase extraction. A reversed phase, Spherisorb RP-8 column and tetrahydrofuran: water:formic acid (42∶56∶2) mobile phase was used for separation. Detection method was by a sensitive fluorimetric method and quantitation was at 395 and 495 nm. The various parameters such as reaction conditions between anilines and fluram, solid phase extraction and chromatographic separation were optimized. Calibrations were linear over the range considered with excellent correlation coefficients (r〉0.999). Relative standard deviations are less than 2.5% and detection limits for aniline,p-toluidine, 4-chloroaniline and 4-bromoaniline were 6, 30, 6 and 8 ng L−1, respectively. This method has been used successfully for the determination of anilines in environmental waters.
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  • 82
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Nauclea diderrichii (de Wild) Merr ; Saponins ; Alkaloid glycoside ; Chrysanthelline A
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A reversed-phase HPLC determination of 11 saponins and an alkaloid glycoside inNauclea diderrichii bark is described. Quantitative analysis of quinovic acid 3-O-α-L-rhamnopyranoside which is one of the main saponins was determined, using Chrysanthelline A as internal standard. The complete separation was achieved in 47 minutes. The method of quantification was validated.
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  • 83
    ISSN: 1612-1112
    Keywords: Gas chromatography ; Column liquid chromatography ; Polychlorinated biphenyls ; Cod liver oil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary By application of a chromatographic column filled with Supelclean ENVI-Carb/Celite and elution with three solvents of different polarity three PCB fractions were obtained. Fraction A contained poly-ortho PCBs, Fraction B mono-ortho PCBs, and Fraction C non-ortho PCBs. The Supelclean ENVI-Carb/Celite column was used in combination with a sample preparation procedure for pre-cleaning of acid-stable chlorinated hydrocarbons such as DDT and its metabolites, HCH isomers, and regulation-relevant PCB congeners. The method was optimized using standard solutions of 55 PCB congeners, 8 chlorinated pesticides and contaminated cod liver oil samples. The influence of traces of remaining matrix on the elution profile of the organochlorine compounds on Supelclean ENVI-Carb/Celite was observed. Quantitation was carried out by GC-ECD with fused silica capillary columns of different polarity.
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  • 84
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Capillary electrophoresis ; Enantiomer separation ; Trimetoquinol ; Denopamine ; Timepidium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Two methods for the direct separation of the enantiomers of trimetoquinol hydrochloride, denopamine and timepidium bromide were investigated, namely capillary zone electrophoresis (CZE) using cyclodextrins (CDs) (CD-CZE) and high-performance liquid chromatography (HPLC) using chiral stationary phases (CSPs). By CZE, separation of the enantiomers of trimetoquinol was successfully achieved when using β-cyclodextrin (β-CD), heptakis(2,6-di-0-methyl)-β-cyclodextrin (DM-β-CD) and β-CD polymer, but direct HPLC separation of the enantiomers was not successful. The enantiomers of denopamine were successfully separated by both CZE, employing DM-β-CD under acidic conditions, and HPLC, with cyclodextrin immobilized CSPs (CD-CSPs). Timepidium bromide, which is a racemic drug, was successfully enantioseparated by both CD-CZE and HPLC with CD-CSPs. The CD-CZE mode was selected as the optical purity testing method for trimetoquinol and denopamine because of the superiority of capillary electrophoretic (CE) techniques. Method validation results obtained clearly show that the CE method would be useful as an quality control method for assessing the optical purity of the drugs.
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  • 85
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Microbore columns ; Column packing ; Slurry viscosity ; Surfactants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The kinematic viscosity of the slurry used for the packing of microbore columns for liquid chromatography has been correlated with the packing efficiency. The columns were packed into polyetheretherketone (PEEK) tubing with an internal diameter of 0.5 mm using unbonded and bonded spherical silica gels under constant pressure conditions. Higher efficiencies were obtained using slurries suspended in aqueous solutions of surfactants, such as polyoxyethylene sorbitane monolaurte (Tween-20) and sodium dodecyl sulphate, in comparison with organic solvents usually used for column packing. The concentration dependencies of the relative kinematic viscosities of six slurries were compared using capillary viscometry. The slurries with low relative viscosities possessed rheological properties which enabled high packing densities, typical of a stable dense random packing arrangement, to be obtained at moderate packing pressures. The unusually low relative viscosities are discussed in terms of a “porous hydrodynamic bearing” model. The techniques used should also be generally applicable to the slurry packing of conventional columns.
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  • 86
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Preservatives-antioxidants ; Eluent pH ; Retention and separation factors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary In this paper the dependence of eluent pH on the efficiency of separation in RP-HPLC is described. Preservatives-antioxidants, esters ofp-hydroxybenzoic acid (methyl, ethyl, propyl and butyl parabens), sorbic acid, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), propyl gallate (PG) and L-ascorbic acid (Vitamine C) were investigated by reversed-phase high-performance liquid chromatography, utilising isocratic elution with UV detection. The mobile phase composition (methanol-water v/v containing 1% acetic acid), was varied in order to obtain the best separation. For all substances analysed the pH, pKa and the fitted pKa values were calculated in different mixtures of mobile phases using the graphical method of Charlot and Trémillon [1]. The retention factors (k) and the separation factors (α) were also calculated.
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  • 87
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    Chromatographia 40 (1995), S. 680-683 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Chiral stationary phase ; Preparative chromatography ; Benzodiazepines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary High performance liquid chromatography chiral stationary phases based on immobilized proteins are well known and commonly used. This work reports on an evaluation of a Human Serum Albumin stationary phase in the preparative separation of benzodiazepines.
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  • 88
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Isoniazid in milk ; SPE preconcentration ; Diode array UV spectra
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A simple, fast, accurate and highly sensitive (5 ng/ml) method for the detection of sionicotinic acid hydrazide (isoniazid) in cattle milk is described, based upon an RP-HPLC system, equipped with a C18 column and a UV detector. Milk was deproteinized with trichloroacetic acid and treated with cinnamaldehyde, before being injected into the HPLC system. Preconcentration of samples on an SPE phenyl cartridge allows the sensitivity of the method to be increased to 0.05 ng/ml using a UV detector. The presence of isoniazid can be confirmed at concentrations as low as 0.4 ng/ml, through a diode-array UV spectrum of isoniazid hydrazone. The analytical procedure here described is routinely applied to investigate the illegal administration of isoniazid to cattle, as it provides a means of screening that can be carried out in a very short time.
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  • 89
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Micellar liquid phases ; Hybrid mobile phases ; Retention behaviour
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Two methods for the description of the retention behaviour of solutes in micellar liquid chromatography are compared. One of them divides the parameter space into triangular subspaces, fitting a different equation in each subspace. The second method makes use of a unique equation, valid in the whole parameter space. In both cases, equations of the type log k=f (μ, ϕ), and 1/k=f (μ, ϕ), (μ and ϕ are the concentration of surfactant and alcohol, respectively), were used to describe the retention. The use of the hyperbolic function, 1/k=c0+c1μ+c3μϕ, to describe the whole parameter space yielded the best prediction. When a small portion of the parameter space was modelled, a simpler hyperbolic function, lacking the μϕ term often led to good results. The logarithmic equations usually gave the poorer predictions.
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  • 90
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; High speed LC ; Pharmaceutical drugs ; Routine analysis ; Fast HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary High performance liquid chromatography (HPLC) is widely used in drug development and quality control. The main objective is to obtain dependable results in the shortest time in order to reduce the time taken to develop drugs and to ensure better quality of the final compound. High speed liquid chromatography (HSLC) helps us to reach this objective. In this paper pratical aspects of HSLC are discussed mainly in terms of instrument requirements and the practical needs of users. Comparison between HPLC and HSLC separations of pharmaceutical preparations are given and discussed. The contribution of HSLC to decrease solvent consumption and consequently analysis cost and its role in reducing environmental pollution, are also mentioned.
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  • 91
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Indirect photometric detection ; Anthraquinone-disulfonate ion ; Inorganic anions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Disodium salts of several anthraquinone-disulfonic acids, which possess large molar absorptivity, were employed as the visualization agent for microcolumn ion chromatography with indirect photometric detection. Detection limits of inorganic anions were improved, e.g., concentration detection limits of chloride, nitrate and sulfate were 0.91–1.6 μmol L−1 at S/N=3, corresponding to mass detection limits of 18–32 fmol. The present system was applied to the determination of inorganic anions in tap water.
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  • 92
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    Chromatographia 40 (1995), S. 404-410 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Solid-phase extraction ; Wine ; Biogenic amines ; Phthalaldehyde
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A suitable method for the determination of 16 biogenic amines in wine has been developed. The method involves clean-up of wine samples using ion-exchange cartridges and a preconcentration step, under controlled vacuum, before derivatization of the amines by treatment with phthalaldehyde (PA) and reversedphase HPLC with gradient elution and fluorimetric detection. Linearity of response was obtained for all the biogenic amines from 100 μg L−1 to mg L−1. Limits of detection for the amines were similar for all PA-derivatives (25–50 μg L−1) and the quantitation limits were about 0.1 mg L−1. After clean-up and preconcentration, the concentration levels increased 10-fold for all amines except putrescine and cadaverine, which gave poor recovery by this method unlike the rest which gave recoveries of almost 90%. The overall process was successfully applied to identify and quantify biogenic amines in several red wines from the Tarragona region.
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  • 93
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Mixed-bed stationary phase ; Anion and cation exchange resins ; Retention mechanism of anions and cations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The retention mechanism of anions and cations on a mixed-bed column packed with a mixture of strong anion and cation exchange resins has been studied. Organic acids and inorganic anions have showed the same elution behaviour on the mixed-bed column as on a single anion exchange column. In contrast, different elution behaviours for cations have been observed between the mixed-bed column and a single cation exchange column. On the mixed-bed column, cations would be eluted by a mixed retention mechanism. The primary retention mechanism is ion exchange with eluent cation (H+) for all cations. The ion exchange mechanism on the sites occupied by anion exchange groups makes a greater contribution for monovalent cations. A partition mechanism between the stationary phase and mobile phase contributes to the separation of Mg2+ and Ca2+ by forming neutral compounds.
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  • 94
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Open tubular columns ; Cross-linked polyacrylate stationary phases ; Photopolymerization ; PTH-amino acids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Fused silica capillaries with an internal diameter of 5 μm were coated with polyacrylate films containing either alkyl or ether functionalities. Polymer networks were created byin situ photopolymerization of acrylic monomers. Capillaries with phase ratios up to 1.3, having excellent separation efficiencies, could be prepared. The performance of these capillaries was demonstrated by the separation of various solute mixtures. Nineteen phenylthiohydantoin (PTH) amino acid derivatives could be separated in less then 10 minutes when using UV detection. Laser Induced Fluorescence detection was used for the separation of polycyclic aromatic hydrocarbons, dansyl (Dns) derivatized amino acids ando-phthalaldehyde (OPA) derivatized aliphatic amines which could be separated with approximately 310,000 plates in less than 3.5 minutes corresponding to around 1500 plates per second.
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  • 95
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    Chromatographia 41 (1995), S. 178-182 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Solid-phase extraction ; Triazine herbicides ; Milk
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary High-performance liquid chromatography with UV detection was used to determine eight triazine herbicides in milk. Solid-phase extraction was performed using a double trap; first, a nonspecific adsorbent (Carbograph), and then a cation exchanger (SCX). Eluate from the SCX was evaporated to dryness under reduced pressure and redissolved in mobile phase. An aliquot was injected into the chromatography, which was operated isocratically in the reverse-phase mode with UV detection at 225 nm. Analytical recoveries for the eight triazines ranged from 73.0% to 92.4%. The limit of sensitivity of this method was about 0.09 ng mL−1 of milk. The method was validated and evaluated by comparison with a method reported in literature.
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  • 96
    ISSN: 1612-1112
    Keywords: Supercritical fluid extraction ; Column liquid chromatography ; Aromatic compounds on silica
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary An SFE-HPLC system has been tested with a simple model aromatic mixture on spiked silica. The spiked silica was extracted with pure CO2 and the extract retained in a trap packed with C18 material. Quantitative separation and reproducity have been investigated and are reported.
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  • 97
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    Chromatographia 41 (1995), S. 445-449 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ion chromatography ; Alkylsulphonic acids ; Conductivity detection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary An ion chromatographic (IC) method has been developed for the separation of alkylsulphonic acid. Two different stationary phases, silica-based and polymer-based ion-exchange resins, were studied using pure ion exchange and/or hydrophobic interaction mechanisms. Correlations between analyte hydrophobicity and eluent polarity were made in order to investigate the possibility of changing the dominant separation mechanism by varying the eluent composition. The alkyl chain lengths of the sulphonic acids analysed ranged from C1 to C9. Detection limits in the submicromolar range were obtained by suppressed conductivity detection.
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  • 98
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Polymeric bonded phase ; Anion-exchange chromatography ; Pulsed amperometric detection ; Carbohydrates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary This paper reports the results of a study on the use of a new polymer-based, strong anion-exchange, stationary phase for rapid and selective separation of carbohydrates and related compounds by high-pH, anion-exchange chromatography with pulsed amperometric detection. The new adsorbent has been obtained by direct nitration of 2.8 μm, spherical non-porous highly crosslinked, styrene-divinylbenzene copolymer beads, followed by reduction of superficially introduced nitro groups with nascent hydrogen and quaternization of the resultant amino groups with iodomethane. It is reported that by optimizing the ionic strength of the mobile phase, columns packed with the new anion-exchanger can be successfully employed to separate, either in isocratic or gradient elution mode, oligosaccharides, positional isomers of gluco-disaccharides, as well as uronic acids and sugar monophosphates.
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  • 99
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Analytical and preparative scales ; Soybean phospholipids ; Mass detector
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary To enable the study of the functional properties of pure soybean phospholipids, a chromatographic method was developed for the preparative fractionation of soybean lecithin. A coarse and irregularly shaped silica gel was used as the stationary phase, whereas the mobile phase consisted of three mixtures of hexane, 2-propanol and water of increasing polarity. These solvents were included in a step gradient, formed by an isocratic pump connected to a solvent changer. To reduce solvent consumption, the procedure was optimised using a method development column. The solvent program and the sample load were shown to affect mainly the purity and recovery of phosphatidylinositol.
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  • 100
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    Chromatographia 41 (1995), S. 521-526 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Phenolic compounds ; Ion-pair solid phase extraction ; On-line trace enrichment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Modifying the most common design for the on-line coupling of a precolumn to reversed phase LC with diode array detection has resulted in reduction of the broadening of the peaks which results when the compounds of interest are strongly retained by a highly hydrophobic sorbent. The modification consists of the desorption of the analytes trapped on the precolumn solely by the organic solvent used to modify the solvent strength of the mobile phase. Results obtained using this design were compared with those obtained with the conventional design, with C18 and PLRP-S precolumns. The performance of the system was also tested with a highly cross-linked styrene-divinylbenzene copolymer (ENVI-chrom P) precolumn for the determination of phenolic compounds in real samples. The advantages and disadvantages are discussed. Ion-pair solid phase extraction is used in order to increase the breakthrough volumes of more polar compounds, mainly phenol. The use of the new design enables phenolic compounds to be determined at the low μg L−1 level with limits of detection ranging between 0.1 and 2 μg L−1 in tap water when a 10 mL sample was analyzed.
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