Operational variables in high-performance liquid chromatography-electrospray ionization mass spectrometry of peptides and proteins using poly(styrene-divinylbenzene) monoliths

Capillary reversed-phase high-performance liquid chromatography (RP-HPLC) utilizing monolithic poly(styrene-divinylbenzene) columns was optimized for the coupling to electrospray ionization mass spectrometry (ESI-MS) by the application of various temperatures and mobile phase additives during peptide and protein analysis. Peak widths at half height improved significantly upon increasing the temperature and ranged from 2.0 to 5.4 s for peptide and protein separations at 70 degrees. Selectivity of peptide elution was significantly modulated by temperature, whereas the effect on proteins was only minor. A comparison of 0.10% formic acid (FA), 0.050% trifluoroacetic acid (TFA), and 0.050% heptafluorobutyric acid (HFBA) as mobile phase additives revealed that highest chromatographic efficiency but poorest mass spectrometric detectabilities were achieved with HFBA. Clusters of HFBA, water, and acetonitrile were observed in the mass spectra at m/z values >500. Although the signal-to-noise ratios for the individual peptides diverged considerably both in the selected ion chromatograms and extracted mass spectra, the average mass spectrometric detectabilities varied only by a factor of less than 1.7 measured with the different additives. Limits of detection for peptides with 500 nl sample volumes injected onto a 60 mm x 0.20 mm monolithic column were in the 0.2-13 fmol range. In the analysis of hydrophobic membrane proteins, HFBA enabled highest separation selectivity at the cost of lower mass spectral quality. The use of 0.050% TFA as mobile phase additive turned out to be the best compromise between chromatographic and mass spectrometric performance in the analysis of peptides and proteins by RP-HPLC-ESI-MS using monolithic separation columns.     

Stereoselective Syntheses of Tetrahydroesterastin-β -Lactam Analogues

A total synthesis of the optically active tetrahydroesterastin β -lactam analogue 2 using Miller's hydroxamate approach is described (Scheme 2). Significant modification of published procedures has resulted in a short and facile stereospecific preparation of the N-[(benzyloxycarbonyl)methyl]-β -lactam 17 starting from the readily available D -serine. This material served as intermediate for the preparation of a variety of N-[(benzyloxycarbonyl)methyl]tetrahydroesterastin β-lactam analogues (Scheme 5).     

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On semianalytic subsets of rigid analytic varieties

We show some topological properties of semianalytic subsets of rigid analytic varieties: curve selection lemma, the closure of a semianalytic subsetS is semianalytic, for every quasi-compact morphismf. As an application we show that a morphismf: X Y of rigid analytic varieties is open at a pointx X if and only if SpecO _X,x SpecO _Y,f(x) is surjective.     

Stimulated emission and laser action of Pr3+-doped YA1O3

Groundstate absorption, fluorescence, excited-state absorption, and stimulated emission of Pr:YA1O₃ were measured in dependence on the polarization. The results reflect the anisotropy of the host lattice. Especially the polarized spectra of the stimulated emission fit very well with the laser data of this material. Seven cw laser transitions in the visible spectral range obtained so far in Pr:YA1O₃ with argon-ion laser pumping were identified in the spectra of the stimulated emission. In addition, two infrared cw laser transitions were realized. The best results were achieved for the³ P ₀³ F ₄ laser transition at 746.9 nm with a maximum slope efficiency of 24.6%, a maximum output power of 49.6 mW, and a laser threshold of 25 mW.     

Efficient Continuous Wave-laser emission of Pr3+-doped fluorides at room temperature

We report, to our knowledge for the first time, Continuous Wave (CW) laser emission at room temperature of Pr:LiYF₄ (Pr.YLF) at six wavelengths: 522, 545, 607, 639.5, 720 and 907.4 nm. The pump source was an argon-ion laser tuned to a wavelength of = 457.9 nm. The maximum output powers at 522 nm (³P_i ³H₅) and 639.5 nm (³P₀ ³F₂) were 144 and 266 mW, respectively. We also observed CW laser action of Pr:GdLiF₄ (Pr: GLF) at = 639 nm and of Pr:KYF₄ (Pr: KYF) at = 642.5 nm.