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121.
Alternative mobile phases for the reversed-phase high-performance liquid chromatography of peptides and proteins 总被引:2,自引:0,他引:2
The use of a high content of acetic acid as mobile phase additive for the reversed-phase high-performance liquid chromatography (RP-HPLC) of several proteins and extracts of biological tissues was evaluated for a divinylbenzene (DVB)-based stationary phase, and the separations obtained with acetic acid gradients in acetonitrile, isopropanol or water were compared with classical polypeptide RP-HPLC on silica C4 with trifluoroacetic acid (TFA)-acetonitrile. The separation patterns for recombinant derived interleukin-1 beta (IL-1 beta) on the C4 column eluted with TFA-acetonitrile and the DVB column eluted with acetic acid-acetonitrile were similar, but only the polymeric column was able to separate the components present in an iodinated IL-1 beta preparation. Neither eluent had any harmful effect on the biological activity of IL-1 beta isolated after RP-HPLC. Several standard proteins could be separated when the polymeric column was eluted with acetic acid gradients in acetonitrile, isopropanol or water and, although the separation efficiency with acetic acid in water was lower than that in combination with classical organic modifiers, insulin, glucagon and human growth hormone (hGH) were eluted as sharp, symmetrical peaks. The recoveries of insulin and hGH were comparable for all three mobile phases (80-90%). The separation patterns obtained from a crude acetic acid extract of a normal and a diabetic, human pancreas analysed using acetic acid gradients with or without organic modifiers were found to be similar and comparable to those obtained on a silica C4 column eluted with an acetonitrile gradient in TFA. The principal differences resulted from the use of different UV wavelengths (215 nm for TFA-acetonitrile, 280 nm for acetic acid). Acetic acid extracts of recombinant derived hGH-producing Escherichia coli were separated on the DVB column eluted with an acetic acid gradient in water. Although the starting material was a highly complex mixture, the hGH isolated after this single-step purification was surprisingly pure (as judged by sodium dodecyl sulphate-polyacrylamide gel electrophoresis). Consequently several (pure) polypeptides and complex biological samples were separated on a polymeric stationary phase eluted with acetic acid gradients in water without the use of organic modifiers. 相似文献
122.
The embedding of attractors and their stable and unstable manifolds can be studied experimentally by controlled addition of chemical species to bring about a particular response. For stable small amplitude oscillations near a Hopf bifurcation from a steady state the embedding can be completely determined even in systems where two of the species are not observable. A quenching of the oscillations by dilution candetermine the steady state concentrations. A species that cannot quench the oscillations almost certainly cannot be an essential component of the oscillation. The method can be extended to a study of attractor associated with subharmonic and quasiperiodic bifurcations and of attractors corresponding to nonperiodic motion. We present preliminary results for a subharmonic bifurcation. 相似文献
123.
K. Mørken 《Constructive Approximation》1991,7(1):195-208
In this paper it is shown how the algebraic product of two spline functions, each represented in terms of B-splines, can again be represented as a linear combination of suitable B-splines. As a corollary to this result we obtain an explicit representation of a given B-spline function in terms of B-splines of some arbitrary higher degree. This generalizes some known results for raising the degree by one. Recurrence relations for both products and degree raising are established that may be useful for computation.Communicated by Larry L. Schumaker. 相似文献
124.
R. Albrecht T. C. Awes P. Beckmann F. Berger D. Bock R. Bock G. Claesson G. Clewing L. Dragon A. Eklund R. L. Ferguson A. Franz S. Garpman R. Glasow H. Å. Gustafsson H. H. Gutbrod G. Hölker J. Idh P. Jacobs K. H. Kampert B. W. Kolb P. Kristiansson H. Löhner I. Lund F. E. Obenshain A. Oskarsson I. Otterlund T. Peitzmann F. Plasil A. M. Poskanzer M. Purschke H. G. Ritter R. Santo S. Saini H. R. Schmidt S. P. Sørensen K. Steffens D. Stüken E. Stenlund M. Tincknell G. R. Young 《Zeitschrift fur Physik C Particles and Fields》1991,51(1):1-10
The production of direct photons has been investigated in reactions ofp and16O projectiles at 60 and 200A·GeV with C and Au nuclei. Photon and 0 spectra have been measured in the pseudorapidity range 1.52.1 for the transverse momentum region 0.4 GeV/cp
T
2.8 GeV/c employing the lead-glass spectrometer SAPHIR. An upper limit of 15% at the 90% confidence level for the direct photon signal relative to the neutral pion production is obtained from the comparison of measured photon spectra with Monte Carlo simulations of the hadronic background based on the reconstructed yield of 0 and mesons. Consequences for a possible phase transition to a quark-gluon plasma are discussed. 相似文献
125.
Erling Størmer 《Inventiones Mathematicae》1992,110(1):63-73
Summary LetL(F
) be theII
1-factor defined by the free groupF
in infinite number of generators. It is shown that for a class of automorphisms ofL(F
) arising from bijections of the set of generators ofF
on itself, and including the free shift, the entropy is zero.Oblatum 15-III-1992 相似文献
126.
Bent Fuglede 《Potential Analysis》1992,1(4):355-371
The first property is a refinement of earlier results of Ch. de la Vallée Poussin, M. Brelot, and A. F. Grishin. Let w=u–v with u, v superharmonic on a suitable harmonic space (for example an open subset of R
n
), and let [w]=[u]–[v] denote the associated Riesz charge. If w0, and if E denotes the set of those points of at which the lim inf of w in thefine topology is 0, then the restriction of [w] to E is 0. Another property states that, if e denotes a polar subset of such that the fine lim inf of |w| at each point of e is finite, then the restriction of [w] to e is 0. 相似文献
127.
We study Schrödinger operators of the form
on
d
, whereA
2 is a strictly positive symmetricd×d matrix andV(x) is a continuous real function which is the Fourier transform of a bounded measure. If
n
are the eigenvalues ofH we show that the theta function
is explicitly expressible in terms of infinite dimensional oscillatory integrals (Feynman path integrals) over the Hilbert space of closed trajectories. We use these explicit expressions to give the asymptotic behaviour of (t) for smallh in terms of classical periodic orbits, thus obtaining a trace formula for the Schrödinger operators. This then yields an asymptotic expansion of the spectrum ofH in terms of the periodic orbits of the corresponding classical mechanical system. These results extend to the physical case the recent work on Poisson and trace formulae for compact manifolds.Partially supported by the USP-Mathematisierung, University of Bielefeld (Forschungsprojekt Unendlich dimensionale Analysis) 相似文献
128.
The molecular structure and conformation of 2,3-dichloro-1-propene have been determined by gas-phase electron diffraction at nozzle temperatures of 24, 90 and 273°C. The molecules exist as a mixture of two conformers with the chlorine atoms anti (torsion angle ∠φ = 0°) or gauche (∠φ = 109°) to each other and with the anti form the more stable. The composition (mole fraction) of the vapor with uncertainties estimated at 2σ was found to be 0.55 (0.08), 0.49 (0.08) and 0.41 (0.10) at 24, 90 and 273°, respectively. These values correspond to an energy difference with estimated standard deviation ΔE° = E°g-E°a = 0.7 ± 0.3 kcal mol?1 and an entropy difference ΔS° = S°g-S°a = 0.6 ± 0.9 cal mol?1 K?1. Some of the diffraction results, together with spectroscopic observations, permit the evaluation of an approximate torsional potential function of the form 2V = V1 (1 - cos φ) + V2 (1 - cos 2φ) + V3 (1 - cos 3φ); the results are V1 = 4.4 ± 0.5, V2 = ?2.9 ± 0.5 and V3 = 4.8 ± 0.2, all in kcal mol?1. The results at 24°C for the distance (ra) and angle (∠α) parameters, with estimated uncertainties of 2σ, are: r(Csp2-H) = 1.098(0.020)Å, r(Csp3-H) = 1.103(0.020)Å, r(CC) = 1.334(0.009)Å, r(C-C) = 1.504(0.013)Å, r(Csp2-Cl) = 1.752(0.021)Å, r(Csp3-Cl) = 1.776(0.020)Å, ∠C-CC = 127.6(1.1)°, ∠Csp3-Csp2-Cl = 110.2(1.0), ∠Csp2-Csp3-Cl = 113.1(1.2)°, ∠H-Csp3-H = 109.5° (assumed), ∠CC-H = 120.0° (assumed) and ∠φ = 108.9(3.4)°. 相似文献
129.
Lutnaes BF Bruås L Kildahl-Andersen G Krane J Liaaen-Jensen S 《Organic & biomolecular chemistry》2003,1(22):4064-4072
The reaction between beta,beta-carotene and BF3-etherates has been investigated, leading to structural elucidation of the blue product, formed in appropriate organic solvents, as a symmetrical charge delocalised dication (lambda(max) 985 nm at room temperature in CHCl3) with considerable stability. The reaction, monitored by EPR studies at -25 degrees C, occurred via free radical intermediates. A C40H56BF3 intermediate was captured by EIMS. The detailed structure of the dication was established by COSY, HSQC, HMBC and 1D and 2D ROESY NMR techniques (600 MHz, CDCl3, -20 degrees C) leading to complete assignments of 1H and 13C chemical shifts and 3J(H,H) coupling constants. The effects of the two delocalised charges on chemical shift (charge distribution) and bond distance (3J(H,H)) were considered. The results are consistent with charge delocalisation mainly in the C-5-C-9 and C-5'-C-9' regions and with bond inversion to retro shifted double bonds in the central C-13-C-13' region. A convention for denoting the charge delocalisation and bond types is presented. The experimental results are discussed relative to previous theoretical calculations of the beta,beta-carotene dication structure. (All-E) and (15-Z)-beta,beta-carotene provided the same dication. The NIR spectra and stability of dications prepared in the same manner from the related carotenes 20,20'-dinor-beta,beta-carotene, heptapreno-beta,beta-carotene and nonapreno-beta,beta-carotene were examined for comparison. Reactions of the beta,beta-carotene dication with selected nucleophiles provided products including isocryptoxanthin, isocarotene and mutatochrome with H2O as nucleophile, and isocryptoxanthin methyl ether, 8-methoxy-7,8-dihydro-beta,beta-carotene and isocarotene with CH3ONa as nucleophile. The formation of these products is rationalised from the structure assigned to the dication. 相似文献
130.
All methionine aminopeptidases exhibit the same conserved metal binding site. The structure of this site with either Co2+ions or Zn2+ions was investigated using density functional theory. The calculations showed that the structure of the site was not influenced by the identity of the metal ions. This was the case for both of the systems studied; one based on the X-ray structure of the human methionine aminopeptidase type 2 (hMetAP-2) and the other based on the X-ray structure of the E. colimethionine aminopeptidase type 1 (eMetAP-1). Another important structural issue is the identity of the bridging oxygen, which is part of either a water molecule or a hydroxide ion. Within the site of hMetAP-2 the results strongly indicate that a hydroxide ion bridges the metal ions. By contrast, the nature of the oxygen bridging the metal ions within the metal binding site of eMetAP-1 cannot be determined based on the results here, due to the similar structural results obtained with a bridging water molecule and a bridging hydroxide ion. 相似文献