Increased Ionization Efficiency Research Articles

Paper-Based Electrochemical Cell Coupled to Mass Spectrometry.

On-line coupling of electrochemistry (EC) to mass spectrometry (MS) is a powerful approach for identifying intermediates and products of EC reactions in situ. In addition, EC transformations have been used to increase ionization efficiency and derivatize analytes prior to MS, improving sensitivity and chemical specificity. Recently, there has been significant interest in developing paper-based electroanalytical devices as they offer convenience, low cost, versatility, and simplicity. This report describes the development of tubular and planar paper-based electrochemical cells (P-EC) coupled to sonic spray ionization (SSI) mass spectrometry (P-EC/SSI-MS). The EC cells are composed of paper sandwiched between two mesh stainless steel electrodes. Analytes and reagents can be added directly to the paper substrate along with electrolyte, or delivered via the SSI microdroplet spray. The EC cells are decoupled from the SSI source, allowing independent control of electrical and chemical parameters. We utilized P-EC/SSI-MS to characterize various EC reactions such as oxidations of cysteine, dopamine, polycyclic aromatic hydrocarbons, and diphenyl sulfide. Our results show that P-EC/SSI-MS has the ability to increase ionization efficiency, to perform online EC transformations, and to capture intermediates of EC reactions with a response time on the order of hundreds of milliseconds. The short response time allowed detection of a deprotonated diphenyl sulfide intermediate, which experimentally confirms a previously proposed mechanism for EC oxidation of diphenyl sulfide to pseudodimer sulfonium ion. This report introduces paper-based EC/MS via development of two device configurations (tubular and planar electrodes), as well as discusses the capabilities, performance, and limitations of the technique.

Capillary Electrophoresis-Electrospray Ionization-Mass Spectrometry for Quantitative Analysis of Glycans Labeled with Multiplex Carbonyl-Reactive Tandem Mass Tags.

Recently developed carbonyl-reactive aminoxy tandem mass tag (aminoxyTMT) reagents enable multiplexed characterization and quantitative comparison of structurally complex glycans between different biological samples. Compared to some previously reported isotopic labeling strategies for glycans, the use of the aminoxyTMT method features a simple labeling procedure, excellent labeling efficiency, and reduced spectral complexity at the MS(1) level. Presence of the tertiary amine functionality in the reporter region of the aminoxyTMT labels leads to increased ionization efficiency of the labeled glycans thus improving electrospray ionization (ESI)-mass spectrometry (MS) detection sensitivity. The use of the labeling reagent also makes electrophoretic separation of the labeled neutral and acidic glycans feasible. In this work, we characterized the ESI and collision induced dissociation (CID) behavior of the aminoxyTMT-labeled neutral and sialylated glycans. For the high-mannose N-glycans and small sialylated oligosaccharides, CID fragmentation of [M + Na + H](2+) provides the most informative MS(2) spectra for both quantitative and qualitative analysis. For complex N-glycans, MS(3) of the protonated Y1(H) ion can be used for relative quantification without interference from the HexNAc fragments. Online capillary electrophoresis (CE)-ESI-MS/MS analyses of multiplexed aminoxyTMT-labeled human milk oligosaccharides (HMOs) and different types of N-glycans released from glycoprotein standards were demonstrated. Improved resolution and quantification accuracy of the labeled HMO isomers was achieved by coupling CE with traveling wave ion mobility (TWIM)-CID-MS/MS. N-Glycans released from human serum protein digests were labeled with six-plex aminoxyTMT and subjected to CE-ESI-MS/pseudo-MS(3) analysis, which demonstrated the potential utility of this glycan relative quantification platform for more complex biological samples.

High-order harmonic generation enhanced by coherent population return

We present computations of high-order harmonic generation in atoms by using the coherent population return technique. This approach consists in two steps: firstly, the active medium is prepared in a coherent superposition of two bound states using a relatively long laser pulse which has a photon energy close to the energy gap between the states and then, the system is driven by an ultrashort infrared laser pulse. We demonstrate, by employing the numerical solution of the time-dependent Schrodinger equation in one and three dimensions, that using this scheme it is possible to enhance the harmonic conversion efficiency by several orders of magnitude. Our numerical results show that, on one hand the increase in ionization efficiency is responsible of an enhancement in the harmonic signal and on the other hand, the coherent preparation of the medium produces a much richer structure of the harmonic emission.

Ionization of solutions using mechanical spray in collison nebulizer: A new simple ion source for mass-spectrometric analysis

Ionization methods for mass-spectrometric analysis based on mechanical liquid spray by a high-speed gas flow (like Sonic Spray Ionization (SSI)) offer some advantages over other ionization methods. Among these one should mention the absence of electric voltage, radioactive sources and heaters. In this paper we suggest using the Collison nebulizer for solution ionization via mechanical spray. Research results into the efficiency of the Collison spray ionization of different solvents used in chemical synthesis are submitted. It is shown that an increase in dipole moment of the solvent molecules leads to increased ionization efficiency for all solvents tested, except water. This seems to result from anomaly high surface tension and vaporization enthalpy for water. High concentrations (>10−4g/ml) of both inorganic salts (NaCl, K2SO4) and organic additive (morphine) give a decrease in the total ion current at the Collison nebulizer outlet. This is assumed to be due both to an increased droplet size of aerosol produced upon spraying and a low dipole moment of an ion-solvation shell complex. Mass-spectra of aqueous K2SO4 and morphine (C17H19NO3) were obtained with the Collison nebulizer. These spectra contained non-identified background peaks along with the peaks of K+ and H+C17H19NO3 that were likely to result from the water ionization and presence of uncontrolled impurities. A relationship between the intensity of mass-spectroscopic peaks of the ions of potassium and morphine and the concentration of appropriate aqueous solution was determined. Mass-spectrometric analysis using the Collison spray ionization gave a detection limit for morphine ∼10−8g/ml and a linear response range for protonated molecular ion (H+C17H19NO3 with m/z=286 from 10−8g/ml to 2×10−7g/ml.

Feasibility assessment of a novel selective peptide derivatization strategy for sensitivity enhancement for the liquid chromatography/tandem mass spectrometry bioanalysis of protein therapeutics in serum

Sensitivity is one major challenge limiting the application of liquid chromatography/tandem mass spectrometry (LC/MS/MS) methods for bioanalysis of proteins. A novel selective peptide derivatization (SPD) strategy was proposed to improve assay sensitivity. The main concept of the SPD strategy is to selectively derivatize surrogate peptides of the target protein in the digests, while not derivatizing the abundant background peptides, thereby improving the separation of target peptides during sample extraction and chromatography, and increasing the sensitivity. Additional benefits may help improve sensitivity include (1) increased ionization efficiency; (2) improved fragmentation pattern; and (3) increased sample extraction recovery of target peptides. Feasibility assessment of the SPD strategy was conducted using BMS-986012, a monoclonal antibody, as the model protein, and with malondialdehyde (MDA) to selectively derivatize the arginine-containing surrogate peptide SLIY in tryptic-digested monkey serum samples. The decreased polarity and basicity of the MDA-derivatized peptide SLIY (MDA-SLIY) helped improve its separation from the background peptides during solid-phase extraction (SPE) and chromatography. The recovery of MDA-SLIY was 36.1-44.2%, which was ~3-fold higher than the recovery of peptide SLIY (11.9-16.1%). There was no significant ion suppression for MDA-SLIY. Overall, SPD improved the sensitivity ~5-fold. SPD methodology was then successfully applied to the development of a sensitive LC/MS/MS assay for BMS-986012 in monkey serum. This work demonstrates the feasibility of the SPD strategy for sensitivity enhancement. SPD can provide a simple, cost-efficient, and antibody-free sample preparation approach to improve sensitivity.

Oxidized fatty acid analysis by charge-switch derivatization, selected reaction monitoring, and accurate mass quantitation

A highly sensitive, specific, and robust method for the analysis of oxidized metabolites of linoleic acid (LA), arachidonic acid (AA), and docosahexaenoic acid (DHA) was developed using charge-switch derivatization, liquid chromatography–electrospray ionization tandem mass spectrometry (LC–ESI MS/MS) with selected reaction monitoring (SRM) and quantitation by high mass accuracy analysis of product ions, thereby minimizing interferences from contaminating ions. Charge-switch derivatization of LA, AA, and DHA metabolites with N-(4-aminomethylphenyl)-pyridinium resulted in a 10- to 30-fold increase in ionization efficiency. Improved quantitation was accompanied by decreased false positive interferences through accurate mass measurements of diagnostic product ions during SRM transitions by ratiometric comparisons with stable isotope internal standards. The limits of quantitation were between 0.05 and 6.0pg, with a dynamic range of 3 to 4 orders of magnitude (correlation coefficient r2>0.99). This approach was used to quantitate the levels of representative fatty acid metabolites from wild-type (WT) and iPLA2γ–/– mouse liver identifying the role of iPLA2γ in hepatic lipid second messenger production. Collectively, these results demonstrate the utility of high mass accuracy product ion analysis in conjunction with charge-switch derivatization for the highly specific quantitation of diminutive amounts of LA, AA, and DHA metabolites in biologic systems.

Characterization of Zhundong subbituminous coal by time-of-flight mass spectrometry equipped with atmospheric pressure photoionization ion source

Zhundong subbituminous coal pretreated with aqueous H2O2 was oxidized by aqueous NaOCl and sequentially extracted with diethyl ether (DEE) and ethyl acetate (EA) to afford DEE- and EA-extractable fractions and residue. Both extracts were esterified with CH2N2 and analyzed using high performance liquid Chromatography/mass spectrometry equipped with electrospray ionization (ESI) or atmospheric pressure photoionization (APPI) ion source. Molecular mass of 60% compounds in the esterified extracts ranged between 200 and 600Da and more than 20% of the esterified extracts had molecular mass over 600Da. Toluene and toluene/anisole (vol./vol.=95/5) were used as dopants for APPI. Low- and non-polar compounds in the esterified extracts were well analyzed using APPI/MS. Compared to toluene/anisole mixture, toluene greatly increased ionization efficiency of APPI. More molecular associations were detected by ESI than by APPI. Heteroatoms were identified in most of the associated species.

Toward Single-Cell Analysis by Plume Collimation in Laser Ablation Electrospray Ionization Mass Spectrometry

Ambient ionization methods for mass spectrometry have enabled the in situ and in vivo analysis of biological tissues and cells. When an etched optical fiber is used to deliver laser energy to a sample in laser ablation electrospray ionization (LAESI) mass spectrometry, the analysis of large single cells becomes possible. However, because in this arrangement the ablation plume expands in three dimensions, only a small portion of it is ionized by the electrospray. Here we show that sample ablation within a capillary helps to confine the radial expansion of the plume. Plume collimation, due to the altered expansion dynamics, leads to greater interaction with the electrospray plume resulting in increased ionization efficiency, reduced limit of detection (by a factor of ~13, reaching 600 amol for verapamil), and extended dynamic range (6 orders of magnitude) compared to conventional LAESI. This enhanced sensitivity enables the analysis of a range of metabolites from small cell populations and single cells in the ambient environment. This technique has the potential to be integrated with flow cytometry for high-throughput metabolite analysis of sorted cells.

Simple and sensitive assay for quantification of oseltamivir and its active metabolite oseltamivir carboxylate in human plasma using high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry: Improved applicability to pharmacokinetic study

Although liquid chromatography/electrospray ionization tandem mass spectrometry-based assays have been reported for the measurement of the antiviral oseltamivir (OS) in human samples, these assays either involve complicated sample pretreatment or lack sensitivity. Here we introduce a straightforward approach to improve the assay performance for OS and its metabolite oseltamivir carboxylate (OSC) in human plasma. A very low concentration of mobile phase modifier can improve the ionization efficiency of both analytes, thus enabling a high sensitivity without any matrix effect. The fast LC gradient further increases the sensitivity by narrowing the peak width (6–9s) and eluting the analytes at higher organic content. The increased ionization efficiency and minimized matrix effects enabled us to introduce a one-step protein precipitation for sample clean-up without compromising the sensitivity. The lower limit of quantification was 0.34ng/mL for both analytes, which was at least 3 times more sensitive than published assays that involve complicated sample pretreatment. The assay involves measurement of analytes and their stable-isotope internal standards in small-volume (30-μL) plasma. Sodium fluoride was utilized to prevent the hydrolysis of OS during and after sampling. The calibration curve was linear over the range of 0.34–1000ng/mL. Accuracy was 95–110% and the precision was 2.2–11.0%. This method was applied successfully to the human pharmacokinetic study of OS, and can estimate the relevant pharmacokinetic parameters of OS with more accuracy. The approach utilized in the optimization of assay performance can be extended to the measurement of other drugs in biomatrices.

Analysis of poly(ethylene glycol) using matrix-assisted laser desorption/ionization-mass spectrometry: Novel matrix systems using ultraviolet light absorber and stabilizer

UV absorbers of orthohydroxyphenyl benzotriazole derivatives and a UV stabilizer (Tinuvin 770) as the matrix system were applied to analysis of poly(ethylene glycol) (PEG) using matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) and investigated. An N2 laser (337nm) was used as the light source. Various UV absorbers of Tinuvin 234, Tinuvin 326, Tinuvin 327, Tinuvin 328, and Tinuvin 329 were used as the matrix. The analytical results were compared with the results of using 2,5-dihydroxybenzoic acid (DHB), the most common matrix for MALDI-MS analysis of PEG. The MALDI-MS spectrum of PEG obtained using Tinuvin 328 or Tinuvin 329 showed higher ionization efficiency than that obtained using DHB. However, by adding Tinuvin 770 to DHB, the ionization efficiency notably enhanced. In contrast to this, for the UV absorbers, the ionization efficiencies were strikingly reduced by adding Tinuvin 770. The increased ionization efficiency of the co-matrix system of DHB+Tinuvin 770 was explained by the formation of the complex of DHB with Tinuvin 770, which improved flexibility of the matrix and enhanced compatibility with PEG. Formation of the complex of the UV absorber with Tinuvin 770 blocked the alcohol group of the UV absorber to remarkably reduce the intermolecular interactions between the matrix and PEG.

Wave modeling in a cylindrical non-uniform helicon discharge

A radio frequency field solver based on Maxwell's equations and a cold plasma dielectric tensor is employed to describe wave phenomena observed in a cylindrical non-uniform helicon discharge. The experiment is carried out on a recently built linear plasma-material interaction machine: The magnetized plasma interaction experiment [Blackwell et al., Plasma Sources Sci. Technol. (submitted)], in which both plasma density and static magnetic field are functions of axial position. The field strength increases by a factor of 15 from source to target plate, and the plasma density and electron temperature are radially non-uniform. With an enhancement factor of 9.5 to the electron-ion Coulomb collision frequency, a 12% reduction in the antenna radius, and the same other conditions as employed in the experiment, the solver produces axial and radial profiles of wave amplitude and phase that are consistent with measurements. A numerical study on the effects of axial gradient in plasma density and static magnetic field on wave propagations is performed, revealing that the helicon wave has weaker attenuation away from the antenna in a focused field compared to a uniform field. This may be consistent with observations of increased ionization efficiency and plasma production in a non-uniform field. We find that the relationship between plasma density, static magnetic field strength, and axial wavelength agrees well with a simple theory developed previously. A numerical scan of the enhancement factor to the electron-ion Coulomb collision frequency from 1 to 15 shows that the wave amplitude is lowered and the power deposited into the core plasma decreases as the enhancement factor increases, possibly due to the stronger edge heating for higher collision frequencies.

The Hydrogen–Deuterium Exchange at α-Carbon Atom inN,N,N-Trialkylglycine Residue: ESI-MS Studies

Derivatization of peptides as quaternary ammonium salts (QAS) is a known method for sensitive detection by electrospray ionization tandem mass spectrometry. Hydrogens at α-carbon atom in N,N,N-trialkylglycine residue can be easily exchanged by deuterons. The exchange reaction is base-catalyzed and is dramatically slow at lower pH. Introduced deuterons are stable in acidic aqueous solution and are not back-exchanged during LC-MS analysis. Increased ionization efficiency, provided by the fixed positive charge on QAS group, as well as the deuterium labeling, enables the analysis of trace amounts of peptides.Electronic supplementary materialThe online version of this article (doi:10.1007/s13361-012-0359-1) contains supplementary material, which is available to authorized users.

Mass spectrometric analysis of phospholipids with phosphate monoester residue using Phos-tag

Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) are growth factor-like bioactive lipids having a phosphate monoester residue. Phos-tag can bind to them and be used both for purification and quantification. In a two-phase solvent system consisting of chloroform/methanol/water, addition of Phos-tag move LPA and S1P from a hydrophilic phase to a hydrophobic phase in the form of their Phos-tag complexes. Using this property, we developed a method for purification of LPA and S1P in biological materials by the phase separation technique. Advantages of use of Phos-tag for detection of LPA and S1P in MALDI-TOF MS are an increase in ionization efficiency and detection as a single-ion form. Homologues of LPA and S1P in natural samples can be quantified by MALDI-TOF MS by using internal standards.

Simultaneous determination of perfluoroalkyl phosphonates, carboxylates, and sulfonates in drinking water

A trace analytical method based on high performance liquid chromatography coupled to quadrupole time-of-flight high resolution mass spectrometry was developed for simultaneous determination of perfluoroalkyl phosphonates (PFPAs, carbon chain lengths C6,8,10), perfluoroalkyl carboxylates (PFCAs, C5–12), and perfluoroalkyl sulfonates (PFSAs, C4,6,8,10) in drinking water (tap water). Analytes were enriched on a mixed mode co-polymeric sorbent (C8 + quaternary amine) using solid phase extraction. Chromatographic separation was achieved on a Zorbax Extend C18 reversed phase column using a mobile phase gradient consisting of water, methanol, and acetonitrile containing 2 mM ammonium acetate and 5 mM 1-methyl piperidine. The mass spectrometer was operated in electrospray negative ion mode. Use of 1-methyl piperidine in the mobile phase resulted in a significant increase in instrument sensitivity for PFPAs through improved chromatographic resolution, background suppression, and increased ionization efficiency. Method detection limits for extraction of 500 mL tap water were in the ranges of 0.095–0.17 ng/L, 0.027–0.17 ng/L, and 0.014–0.052 ng/L for PFPAs, PFCAs, and PFSAs, respectively. Whole method recoveries at a spiking level of 0.5 ng/L to 500 mL HPLC grade water were 40–56%, 56–97%, and 55–77% for PFPAs, PFCAs, and PFSAs, respectively. A matrix effect (signal enhancement) was observed in the detection of PFPAs in tap water extracts, leading to calculated recoveries of 249–297% at a 0.5 ng/L spiking level. This effect resulted in an additional improvement of method sensitivity for PFPAs. To compensate for the matrix effect, PFPAs in tap water were quantified using matrix-matched and extracted calibration standards. The method was successfully applied to the analysis of drinking water collected from six European countries. PFPAs were not detected except for perfluorooctyl phosphonate (PFOPA) at close to the detection limit of 0.095 ng/L in two water samples from Amsterdam, the Netherlands. Highest levels were found for perfluorobutane sulfonate (PFBS, 18.8 ng/L) and perfluorooctanoate (PFOA, 8.6 ng/L) in samples from Amsterdam as well as for perfluorooctane sulfonate (PFOS, 8.8 ng/L) in tap water from Stockholm, Sweden.

Identification of a Gene Essential for the First Committed Step in the Biosynthesis of Bacteriochlorophyll c

Bacteriochlorophylls (BChls) c, d, and e are the major chlorophylls in chlorosomes, which are the largest and one of the most efficient antennae produced by chlorophototrophic organisms. In the biosynthesis of these three BChls, a C-13(2)-methylcarboxyl group found in all other chlorophylls (Chls) must be removed. This reaction is postulated to be the first committed step in the synthesis of these BChls. Analyses of gene neighborhoods of (B)Chl biosynthesis genes and distribution patterns in organisms producing chlorosomes helped to identify a gene (bciC) that appeared to be a good candidate to produce the enzyme involved in this biochemical reaction. To confirm that this was the case, a deletion mutant of an open reading frame orthologous to bciC, CT1077, was constructed in Chlorobaculum tepidum, a genetically tractible green sulfur bacterium. The CT1077 deletion mutant was unable to synthesize BChl c but still synthesized BChl a and Chl a. The deletion mutant accumulated large amounts of various (bacterio)pheophorbides, all of which still had C-13(2)-methylcarboxyl groups. A C. tepidum strain in which CT1077 was replaced by an orthologous gene, Cabther_B0081 [corrected] from "Candidatus Chloracidobacterium thermophilum" was constructed. Although the product of Cabther_B0081 [corrected] was only 28% identical to the product of CT1077, this strain synthesized BChl c, BChl a, and Chl a in amounts similar to wild-type C. tepidum cells. To indicate their roles in the first committed step of BChl c, d, and e biosynthesis, open reading frames CT1077 and Cabther_B0081 [corrected] have been redesignated bciC. The potential mechanism by which BciC removes the C-13(2)-methylcarboxyl moiety of chlorophyllide a is discussed.

3‐Mercaptopropionic acid modified ZnSe quantum dots as the matrix for direct surface‐assisted laser desorption/ionization mass spectrometric analysis of peptides/proteins from sodium salt solution

This study describes a strategy of using zinc selenium quantum dots (ZnSe QDs) modified with 3-mercaptopropionic acid (3-MPA) as the matrix for direct analysis of peptides and proteins from sodium salt solution in surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS). The enhancement of detection sensitivity for these biomolecules was due to the adsorption of positively charged peptides or proteins onto the surfaces of negatively charged ZnSe-3MPA QDs via electrostatic interactions resulting in an increase in ionization efficiency for sodium adduct ions ([M+Na](+)). The applicability of the current approach was demonstrated for a variety of peptides, including leucine-enkephalin, methione-enkephalin, HW6, substance P and angiotensin II, and proteins (cytochrome c, myoglobin and lysozyme). Signal intensities of these peptides or proteins can be enhanced by 25-95 times compared with those obtained by LDI-MS in the absence of ZnSe-3MPA QDs. Applying ZnSe-3MPA QDs to serve as the matrix in SALDI-MS is a simple and effective approach for direct analysis of peptide and protein molecules from sodium salt solution without any pretreatment as the peptides and proteins can be successfully detected as sodium adduct ions ([M+Na](+)).

Alkylating tryptic peptides to enhance electrospray ionization mass spectrometry analysis.

A major limitation of mass spectrometry-based proteomics is inefficient and differential ionization during electrospray ionization (ESI). This leads to problems such as increased limits of detection and incomplete sequence coverage of proteins. Incomplete sequence coverage is especially problematic for analyses that require the detection and identification of specific peptides from a protein, such as the analysis of post-translational modifications. We describe here the development and use of aldehyde-based chemistry for the alkylation of peptide primary amines to increase peptide hydrophobicity, providing increased ionization efficiency and concomitant signal enhancement. When employed to modify the peptide products of protein tryptic digests, increased sequence coverage is obtained from combined modified and unmodified digests. To evaluate the utility of alkylation of peptides for selected reaction monitoring (SRM) assays, we alkylated a peptide from the protein Oct4, known to play a role in regulating stem cell differentiation. Increased chromatographic retention and ionization efficiency is observed for the alkylated Oct4 peptide compared to its unmodified form.

Suitability of tetrahydofuran as a dopant and the comparison to other existing dopants in dopant‐assisted atmospheric pressure photoionization mass spectrometry in support of drug discovery

In this paper, we investigated the suitability of tetrahydofuran (THF) as a dopant and compared it against other common dopants for atmospheric pressure photoionization mass spectrometry (APPI-MS). In a systematic analysis of 37 drug standards and 100 Wyeth proprietary drug candidates, THF was found to increase ionization efficiency as high as 33-fold when introduced through a syringe pump at a flow rate of 20 microL/min, and as high as 114-fold when introduced through the mobile phase at 100 microL/min. As a dopant, THF is as effective as acetone, better than anisole, and slightly less effective than toluene for the majority of the test compounds. The increase in ionization efficiency by THF was found to be compound-dependent. THF was more effective in facilitating the ionization of polar compounds than of non-polar compounds. With THF, toluene and acetone as dopants, a single type of molecular ion ([M+H](+) or M(+*)) is produced for analyte molecules. However, anisole can cause the formation of an ion cluster for polar analytes. The cluster contains [M-2H+H](+), M(+*), and [M+H](+) ions with varied ratios. This complexity may make interpretation of spectra difficult for unknown compounds when complimentary data are not available. Our findings indicate that THF is a suitable dopant in the daily usage for increasing ionization efficiency, especially when THF is used as the mobile phase or as an organic modifier in the mobile phase.

THE UDF05 FOLLOW-UP OF THE HUBBLE ULTRA DEEP FIELD. II. CONSTRAINTS ON REIONIZATION FROMZ-DROPOUT GALAXIES

We detect three (plus one less certain) z850-dropout sources in two separate fields (Hubble Ultra Deep Field and NICP34) of our UDF05 Hubble Space Telescope Near-Infrared Camera and Multi-Object Spectrometer images. These z ∼ 7 Lyman-break Galaxy (LBG) candidates allow us to constrain the Luminosity Function (LF) of the star-forming galaxy population at those epochs. By assuming a change in only M∗ and adopting a linear evolution in redshift, anchored to the measured values at z ∼ 6, the best-fit evolution coefficient is found to be 0.43 ± 0.19 mag per unit redshift (0.36 ± 0.18, if including all four candidates), which provides a value of M∗(z = 7.2) =− 19.7 ± 0.3. This implies a drop in the luminosity density in LBGs by a factor of ∼ 2–2.5 over the ∼ 170 Myr that separate z ∼ 6 and z ∼ 7, and a steady evolution for the LBG LF out to z ∼ 7, at the same rate that is observed throughout the z ∼ 3–6 period. This puts a strong constraint on the star-formation histories of z ∼ 6 galaxies, whose ensemble star-formation rate (SFR) density must be lower by a factor of 2 at ∼ 170 Myr before the epoch at which they are observed. In particular, a large fraction of stars in the z ∼ 6 LBG population must form at redshifts well above z ∼ 7. The rate of ionizing photons produced by the LBG population consistently decreases with the decrease in the cosmic SFR density. Extrapolating this steady evolution of the LF out to higher redshifts, we estimate that galaxies would be able to reionize the universe by z ∼ 6, provided that the faint-end slope of the z> 7 LF steepens to α ∼− 1.9 and that faint galaxies with luminosities below the current detection limits contribute a substantial fraction of the required ionizing photons. This scenario, however, gives an integrated optical depth to electron scattering that is ∼ 2σ below the Wilkinson Microwave Anisotropy Probe-5 measurement. Therefore, altogether, our results indicate that, should galaxies be the primary contributors to reionization, either the currently detected evolution of the galaxy population slows down at z 7, or the LF evolution must be compensated by a decrease in metallicity and a corresponding increase in ionization efficiency at these early epochs.

Diethylation Labeling Combined with UPLC/MS/MS for Simultaneous Determination of a Panel of Monoamine Neurotransmitters in Rat Prefrontal Cortex Microdialysates

The primary challenge associated with the development of an LC/MS/MS-based assay for simultaneous determination of biogenic monoamine neurotransmitters such as norepinephrine (NE), dopamine (DA), serotonin (5-HT), and normetanephrine (NM) in rat brain microdialysates is to improve detection sensitivity. In this work, a UPLC/ MS/MS-based method combined with a diethyl labeling technique was developed for simultaneous determination of a panel of monoamines in rat prefrontal cortex microdialysates. The chromatographic run time is 3.5 min/ sample. The limits of detection of the UPLC/MS/MS-based method for NE, DA, 5-HT/ and NM, with/without diethyl labeling of monoamines, are 0.005/0.4 (30/2367 pM), 0.005/0.1 (33/653 pM), 0.005/0.2 (28/1136 pM), and 0.002/0.2 ng/mL (11/1092 pM), respectively. Diethyl labeling of amino groups of monoamines affords 20-100 times increased detection sensitivity of corresponding native monoamines during the UPLC/MS/MS analysis. This could result from the following: (1) improved fragmentation patterns; (2) increased hydrophobicity and concomitantly increased ionization efficiency in ESI MS and MS/MS analysis; (3) reduced matrix interference. This labeling reaction employs a commercially available reagent, acetaldehyde-d4, to label the amine groups on the monoamines via reductive amination. It is also simple, fast (approximately 25-min reaction time), specific, and quantitative under mild reaction conditions. Data are also presented from the application of this assay to monitor the drug-induced changes of monoamine concentrations in rat prefrontal cortex microdialysate samples followed by administration of SKF 81297, a selective D1 dopamine receptor agonist known to elevate the extracellular level of the neurotransmitters DA and NE in the central nervous system.