A mechanistic study of electrospray mass spectrometry: charge gradients within electrospray droplets and their influence on ion response.

A model has been developed to account qualitatively for the effects of ion pairing, surface activity, and electrophoretic mobility in electrospray mass spectrometry. The model is tested with various salt and amino acid mixtures. The data suggest that the axial charge gradient arising from electrophoretic separation at droplet genesis may persist within the electrosprayed droplets at least until the first droplet fission, accounting for the field dependence of detected ion clustering of quaternary ammonium salts and for the relatively field-invariant charge distribution of horse heart myoglobin samples.

Abeta amyloid fibrils possess a core structure highly resistant to hydrogen exchange.

We describe here experiments designed to characterize the secondary structure of amyloid fibrils of the Alzheimer's amyloid plaque peptide Abeta, using hydrogen-deuterium exchange measurements evaluated by mass spectrometry. The results show that approximately 50% of the amide protons of the polypeptide backbone of Abeta(1-40) resist exchange in aqueous, neutral pH buffer even after more than 1, 000 h of incubation at room temperature. We attribute this extensive, strong protection to H-bonding by residues in core regions of beta-sheet structure within the fibril. The backbone amide hydrogens exchange at variable rates, suggesting different degrees of protection within the fibril. These data suggest that it is unlikely that the entire Abeta sequence is involved in H-bonded secondary structure within the amyloid fibril. Future studies using the methods described here should reveal further details of Abeta fibril structure and assembly. These methods also should be amenable to studies of other amyloid fibrils and protein aggregates.

A symmetry-based formal synthesis of zaragozic acid A.

A symmetry-based strategy for the synthesis of the zaragozic acids is reported. Two enantioselective dihydroxylations were used to establish the absolute configuration of a C(2) symmetric intermediate. Noteworthy transformations include a group-selective lactonization, which accomplished an end-differentiation of a pseudo-C(2) symmetric intermediate. Late stage protecting group adjustments and oxidations accomplished a formal synthesis of zaragozic acid A.

Simultaneous analysis of butene isomer mixtures using process mass spectrometry.

The feasibility of simultaneous analysis of mixtures containing two to four butene isomers and up to six total components using process mass spectrometry is assessed. As for typical (nonisomeric) applications of process mass spectrometry, simultaneous analysis is based on the assumption that the electron ionization mass spectra of mixtures are linear combinations of the spectra of the individual constituents. Limits of detection for binary isomer mixtures are on the order of 0.1% to 10%, limited by the ability to distinguish small differences between similar spectra. As spectral and mixture complexity increase, both accuracy and precision decrease. Not surprisingly the similarity of the spectra of stereoisomers cis- and trans-2-butene is greater than that of the other (nonstereoisomeric) isomer pairs, and mixtures containing both cis- and trans-2-butene are the most difficult to quantitate. However, even for mixtures of all four butenes, accuracy (root-mean-square error = 2.43%), precision (average coefficient of variation = 6.72%), and linearity (correlation coefficient of a plot of measured versus actual concentration r2 = 0.985 +/- 0.002) are reasonably good.

Osteoarthritis of the knee: isokinetic quadriceps exercise versus an educational intervention.

OBJECTIVE: To evaluate the effects of isokinetic exercise versus a program of patient education on pain and function in older persons with knee osteoarthritis. DESIGN: A randomized, comparative clinical trial, with interventions lasting 8 weeks and evaluations of 12 weeks. SETTING: An outpatient Veterans Affairs Medical Center clinic and an affiliated university hospital. PATIENTS: One hundred thirteen men and women between 50 and 80 years old with diagnosed osteoarthritis of the knee; 98 completed the entire assigned treatment. INTERVENTION: Patients received either a regimen of isokinetic exercise of the quadriceps muscle three times weekly over 8 weeks or a series of 4 discussions and lectures led by health care professionals. MAIN OUTCOME MEASURES: Variables studied for change were isokinetic and isometric quadriceps strength, pain and function determined by categorical and visual analog scales, and overall status using physician and patient global evaluations by the Arthritis Impact Scale, version 2, Western Ontario McMaster's Arthritis Index, and Medical Outcome Study Short Form 36. RESULTS: Both treatment groups showed significant strength gains (p < .05), which occurred over a wider velocity spectrum for the exercise group. Exercised patients also had improved pain scores for more of the variables measured than those receiving education. Both groups had positive functional outcomes and slightly improved measures of overall status. CONCLUSIONS: Isokinetic exercise is an effective and well-tolerated treatment for knee osteoarthritis, but a much less costly education program also showed some benefits.

Time-Dependent Permeance of Gas Mixtures through Zeolite Membranes.

The time-dependent permeation behavior of binary gas mixtures through a ZSM-5 zeolite membrane was studied. Although steady-state permeation rates were indistinguishable for CO(2) and N(2) or for cis- and trans-2-butene in binary mixtures, differences in the rate of approach to steady state allowed component distinction. In "normal" systems, one component is initially enriched in the permeate following application of a pulse of analyte gas to the membrane, and then disappears more quickly upon termination of the pulse. Mixtures of cis- and trans-2-butene exhibit qualitatively different behavior; the permeate is enriched in cis-2-butene during both the leading and trailing edges of a sample pulse (though not at steady state). These differences in permeation behavior reflect different balances among multiple transport mechanisms through the zeolite membrane, thought to reflect a combination of selective component sorption and intracrystalline diffusion; in the case of cis- and trans-2-butene, these two factors oppose one another. It is known that this mechanistic complexity can engender synergistic effects, wherein the presence of one component can affect the permeation of another. These may limit applicability to true "unknowns", but resulting complications should be less problematic in well-defined process applications.

Effects of molecular entanglements during electrospray of high molecular weight polymers.

Concentration-dependent bimodal size distributions (comprised of single-molecule particles and multimolecule clusters) observed by microscopic examination of particles collected during electrospray (ES) of dilute solutions of high molecular weight polymers suggest that chain entanglement can interfere with the droplet subdivisions believed to be intrinsic to the electrospray process. The feasibility of such interference is discussed in the context of the spray model of Kebarle, along with its potential impact on the ES mass spectrometry of macromolecules.

Restaurant consumer acceptance of beef loin strip steaks tenderized with calcium chloride.

Beef strip loins from either the right or left side of 22 carcasses of Bos indicus-type steers were injected with 200 mM calcium chloride (CaCl2) solution at 5% (wt/wt) to determine its effect on tenderness and other selected quality traits of steaks. Loins from opposite sides of the carcasses were untreated and served as the control. The steaks were evaluated for tenderness, juiciness, flavor intensity, tenderness acceptability, and overall acceptability by 62 restaurant consumers over a 6-wk period. The CaCl2 injection improved (P < .05) tenderness and flavor intensity ratings by the restaurant consumers. Tenderness acceptability and overall acceptability were improved 23 and 17%, respectively, by the CaCl2 injection. Flavor was not compromised by the CaCl2 injection. The CaCl2-treated steaks were rated superior(P < .05) for flavor compared to the control steaks. Restaurant consumers preferred the beef loin strip steaks injected with 200 mM CaCl2 at 5% (wt/wt). The results of this study are interpreted to indicate that, from a restaurant consumer perspective, CaCl2 injection is an acceptable means of making beef a more consistently tender product.

Durable gold-coated fused silica capillaries for use in electrospray mass spectrometry.

A simple procedure for preparing gold-coated silica capillaries for use in electrospray ionization mass spectrometry is described. The tip of the capillary is mechanically tapered to a fine point, and a thin film of gold is vapor deposited on the outer surface following treatment with an organofunctional silane. The performance characteristics of these durable capillaries as continuous infusion sources are examined, and their utility in on-line capillary electrophoresis mass spectrometry is demonstrated.

Fast-atom bombardment-induced condensation of glycerol with ammonium surfactants II: Time dependence of mass spectra and tandem mass spectra.

Studies of fast-atom bombardment (FAB)-induced condensation between trimethyltetradecylammonium cations and glycerol have been extended to consider spectral time dependence. To enhance reproducibility of time dependence, a modified FAB target was used. FAB mass spectrometry of deuterium-labeled surfactants and FAB/collision-induced dissociation (CID) of nonlabeled material demonstrate that products of condensation at the surfactant "head group" predominate early in the analysis, while tail adducts become prominent later. This time dependence correlates with the expected surface activity of the products. It is incompatible with gas-phase reaction, but consistent with reaction in the condensed phase. Subtle variations in the surface activities of various condensation products (derived from changes in the number of hydroxyls from the reactive glycerol radical or in the position of attack along the surfactant chain) are reflected in the time dependence of FAB and CID spectra. CID spectra of deuterium-labeled cations provide evidence for intramolecular hydrogen transfer from the surfactant tail to the head within a surfactant radical. This transfer shows no significant kinetic isotope effect.

Determination of flux from a saddle field fast-atom bombardment gun.

The flux or beam density (equivalent current/area) of xenon atoms striking the sample target from a saddle field fast-atom bombardment (FAB) gun has been compared with that from a cesium ion gun mounted on the same instrument. A shielded Faraday cup mounted on the end of a solids probe was used to measure directly the flux of the Cs(+) beam. Samples of methylene blue in glycerol solution were then exposed to the ion beam at different fluxes and the extents of reduction were measured. The extent of reduction varied linearly with flux up to a value of about 1.16 × 10(13) particles s(-1) cm(-2) (1.85 µ cm(-2)); above this level, the reduction effect appeared to saturate. FAB spectra were obtained from the same dye solution by using varying settings of the FAB gun. By comparing the extents of reduction of the dye from the two guns, the flux from the atom gun could be estimated. Observation of luminescence from a CsI-coated target allowed estimation of the area of the atom beam. The atom beam "equivalent current" could then be calculated by multiplying the flux times the area. It was noted that for given settings, the flux from the atom gun depended on the physical condition of the gun electrodes. With new electrodes, a flux ≥ 1.16 × 10(13) particles s(-1) cm(-2) was obtained with nominal gun emission currents of 0.60-1.0 mA. Electrodes used extensively, but freshly cleaned, provided a flux of ∼ 8 × 10(12) particles s(-1) cm(-2) at nominal emission currents of 0.40-1.0 mA. With dirty electrodes this flux could only be achieved at the highest (1.0 mA) emission current. This decline in performance occurs over a matter of months as a result of contamination and erosion of the electrodes during use. Such behavior can adversely affect spectral reproducibility even when nominal FAB gun voltage and emission current are carefully reproduced.

Secondary ion emission from solutions: time dependence and surface phenomena.

The temporal behavior of FAB mass spectra from glycerol solutions of tetradecyltrimethylammonium bromide (C14H29-N(CH3)3Br, TTAB) and tetraethylammonium iodide (TEAI) was investigated. FAB spectra of the TTAB solution displayed a continuous decrease in TTA+ with time. Spectra obtained from the TEAI solution were initially invariant for several minutes and then displayed a gradual increase in the relative abundance of TEA+ to a maximum, followed by a precipitous drop in ion intensity. Secondary ion images of droplets of TTAB solution showed that emission of both TTA+ and glycerol secondary ions was homogeneous across the sample. Secondary ion images of droplets of TEAI solution showed heterogeneous and segregated emission of both TEA+ and protonated glycerol. Results from the FAB spectra and the secondary ion images were correlated and rationalized on the basis of surface tension-induced mass transport and matrix evaporation.

m-nitrobenzyl alcohol electrochemistry in fast atom bombardment mass spectrometry.

The efficacy of m-nitrobenzyl alcohol (NBA) as a solvent (matrix) for fast atom bombardment (FAB) mass spectrometry of a group of pyrazolate-bridged dirhodium A-frame complexes has been assessed. Although NBA is frequently used to mitigate the formation of artifacts in FAB/MS of organometallics and other materials susceptible to bombardment-induced reactions, substantial evidence indicates that such reactions cause the formation of artifacts in the spectra obtained here. Parallel absorption spectroscopic studies have established that NBA is capable of inducing both oxidation and reduction reactions independent of ion bombardment, depending on analyte reduction half-wave potential (E1/2). From the known electrochemistry of the complexes studied, it can be estimated that 1020 mV > E1/2 > 500 mV for the reaction of NBA serving as a reducing agent, while 500 mV > E1/2 > 424 mV for the reduction potential of NBA. However, in the presence of bombardment the former E1/2 must be at least as low as 356 mY, and the latter E1/2 must be at least as high as 1188 mY. The kinetics of redox reactions involving NBA, and therefore their influence on the appearance of FAB mass spectra, will be highly sample-dependent. However, this study illustrates an important potential role for redox reactions when NBA is used as a solvent, especially in the presence of bombardment in FAB/MS. Although analyte reaction products could be identified, substantial efforts aimed at identifying NBA oxidation and reduction products did not yield any definitive results due to the complexity of product mixtures.

Fast atom bombardment-induced condensation of glycerol with ammonium surfactants. I: Regioselectivity of the adduct formation.

Fast atom bombardment promotes condensation between trimethyl tetradecyl ammonium cations and the glycerol matrix. Bond formation at both the head and tail of the surfactant is demonstrated by low energy collision-induced dissociation (ClD) of deuterium-labeled precursors, with a preponderance of the reaction apparently occurring at the tail. Two distinct ClD pathways are identified for each kind of adduct (head- and tail-attack). Evidence is presented for the detection of distonic radical cations of the surfactant, complexed (solvated) with glycerol.

Mass spectrometric signature of S-prenylated cysteine peptides.

The fast atom bombardment mass spectra of peptides containing S-prenylated cysteine display signature fragmentations characteristic of this modified amino acid. The fragmentation is independent of the nature of the cysteine carbonyl substituent, easily differentiates prenyl from nonprenyl alkylation, and readily identifies the oligomer count of the prenyl. This screening method, which requires little time, effort, or material (compared with previous analysis methods based on chemical degradation), greatly facilitates the identification of these prenylated proteins.

Charge-remote fragmentation in a hybrid (BEqQ) mass spectrometer to determine isotopic purity in selectively polydeuterated surfactants.

The combination of fast atom bombardment with charge-remote fragmentation using a hybrid (BEqQ) mass spectrometer was used successfully to assess and localize the extent of selective deuterium isotope labeling of tetradecyItrimethylammonium bromides. Spectral details reveal a new reaction that can give rise to ions isobaric with those formed by chargeremote fragmentation.

Improving fast atom bombardment mass spectra: The influence of some controllable parameters on spectral quality.

The relative effects of adjustable fast atom bombardment (FAB) parameters (choice of matrix, primary atom flux, and primary atom energy) on the appearance of FAB spectra (including signal-to-noise, signal-to-background, and signal-to-matrix ratios) of several organic dyes have been investigated. Beam-induced chemical damage is minimized by lowering the primary atom flux, by raising the primary atom energy, and by selecting a matrix with radical scavenging properties (e.g., m-nitrobenzyl alcohol). The relative importance in minimizing this chemical damage is choice of matrix > primary atom flux > (nominal) primary atom energy, but optimization of the parameters involves a trade-off between sensitivity and damage. The effect of these parameters on thermal damage (fragmentation) is much less. It can be concluded from comparison of the dyes that the extent of beam damage does not depend simply on the standard reduction potential of the analyte.