Enhancement of atmospheric pressure chemical ionization for the determination of free and glycine-conjugated bile acids in human serum

A highly sensitive and accurate method based on the precolumn derivatization of bile acids (BA) with a high ionization efficiency labeling reagent 1,2-benzo-3,4-dihydrocarbazole-9-ethyl-benzenesulfonate (BDEBS) coupled with LC/MS has been developed. After derivatization, BA molecules introduced a weak basic nitrogen atom into the molecular core structure that was readily ionized in commonly used acidic HPLC mobile phases. Derivatives were sufficiently stable to be efficiently analyzed by atmospheric pressure chemical ionization (APCI)-MS/MS in positive-ion mode. The MS/MS spectra of BA derivatives showed an intense protonated molecular ion at m/z [M + H](+). The collision-induced dissociation of the molecular ion produced fragment ions at [MH - H2O](+), [MH - 2H(2)O](+), [MH - 3H(2)O](+). The characteristic fragment ions were at m/z 320.8, 262.8, and 243.7 corresponding to a cleavage of N - CO, O - CO, and C - OCC, respectively, and bonds of derivatized molecules. The selected reaction monitoring, based on the m/z [M + H]+ -> [MH - H2O](+), [MH - H2O](+), [MH - 2H(2)O](+), [MH-3H(2)O](+), 320.8, 262.8, and 243.7 transitions, was highly specific for the BA derivatives. The LODs for APCI in a positive-ion mode, at an S/N of 5, were 44.36-153.6 fmol. The validation results showed high accuracy in the range of 93-107% and the mean interday precision for all standards was < 15% at broad linear dynamic ranges (0.0244-25nmol/mL). Good linear responses were observed with coefficients of > 0.9935 in APCI/MS detection. Therefore, the facile BDEBS derivatization coupled with mass spectrometric analysis allowed the development of a highly sensitive and specific method for the quantitation of trace levels of the free and glycine-conjugated BA from human serum samples.

Reconstruction of atomic ionization probabilities in intense laser fields

Application of a parallel-projection inversion technique to z-scan spectra of multiply charged xenon and krypton ions, obtained by non-resonant field ionization of neutral targets, has for the first time permitted the direct observation of intensity-dependent ionization probabilities. These ionization efficiency curves have highlighted the presence of structure in the tunnelling regime, previously unobserved under full-volume techniques.

Process-kinetics in facing targets sputtering of multi-component oxide thin films

The kinetics of the processes in facing targets sputtering of multicomponent oxide films is presented. The novel configuration of the process exhibits an enhanced ionization efficiency. Discharge diagnostics performed using optical emission spectroscopy revealed strong dependence of plasma parameters on process conditions. Numerical simulation based on thermalization and diffusion of sputtered atoms has been performed to estimate the transport efficiency in off-axis mode. Composition, structure and epitaxial quality of YBa2Cu3O7-x films prepared was found to be strongly dependent on atomic flux ratios (of Cu/Y and Ba/Y) arriving at the substrate, resputtering effect and phase stability of YBa2Cu3O7-x These studies have been shown to be useful in understanding the complex processes that occur in sputtering of multicomponent films. (C) 1999 Elsevier Science S.A. All rights reserved.

Spektroskopie und Ultraspurenanalyse von Plutonium mittels Resonanzionisations-Massenspektrometrie

Die Resonanzionisations-Massenspektrometrie (RIMS) ist sowohl für spektroskopische Untersuchungen seltener Isotope als auch für den Ultraspurennachweis langlebiger radioaktiver Elemente einsetzbar. Durch die mehrstufige resonante Anregung atomarer Energieniveaus mit anschließender Ionisation mit Laserlicht wird eine sehr hohe Elementselektivität und Ionisationseffizienz erreicht. Der nachfolgende massenselektive Ionennachweis liefert eine gute Isotopenselektivität zusammen mit einer effektiven Untergrundunterdrückung. Ein wichtiger Bestandteil der RIMS-Apparatur ist ein zuverlässig arbeitendes, leistungsstarkes Lasersystem für die Resonanzionisation. Im Rahmen dieser Arbeit wurde ein von einem hochrepetierenden Nd:YAG-Laser gepumptes, aus drei Titan-Saphir-Lasern bestehendes System fertig aufgebaut und in den Routinebetrieb überführt. Die Titan-Saphir-Laser liefern im Durchstimmbereich von 730 - 880 nm eine mittlere Leistung von bis zu 3 W pro Laser bei einer Linienbreite von 2 - 3 GHz. Sie lassen sich computergesteuert in ihren Wellenlängen durchstimmen. Die mittels Resonanzionisation erzeugten Ionen werden dann in einem Flugzeit-Massenspektrometer entsprechend ihrer Masse aufgetrennt und mit einem Kanalplattendetektor nachgewiesen.Als Voraussetzung für die isotopenselektive Ultraspurenanalyse von Plutonium wurden mit diesem Lasersystem die Isotopieverschiebungen eines effizienten, dreistufigen Anregungsschema für Plutonium bestimmt. Die Laserleistungen reichen zur vielfachen Sättigung der ersten beiden Anregungsschritte und zur zweifachen Sättigung des dritten Anregungsschritts aus.Außerdem wurden die Ionisationsenergien von Pu-239 und Pu-244 zur Untersuchung ihrer Isotopenabhängigkeit bestimmt. Die beiden Ionisationsenergien sind im Rahmen der erreichten Genauigkeit bei einem Meßwert von IP239-IP244 = 0,24(82) cm^-1 gleich.Die Nachweiseffizienz der RIMS-Apparatur für Plutonium wurde in Effizienzmessungen zu 10^-5 bestimmt. Durch die gute Untergrundunterdrückung ergab sich daraus eine Nachweisgrenze von 10^6 Atomen bei der Messung eines Plutoniumisotops. Die Bestimmung der Isotopenverhältnisse von Proben mit einer zertifizierten Isotopenzusammensetzung lieferte eine gute Übereinstimmung der Meßwerte mit den angegebenen Zusammensetzungen.Die RIMS-Apparatur wurde zur Bestimmung des Gehalts und der Isotopenzusammensetzung von Plutonium in Meerwasser- und Staubproben eingesetzt.Auf Grund der Isotopenzusammensetzung konnte gezeigt werden, daß das Plutonium bei den meisten Proben aus dem Fallout von oberirdischen Kernwaffentests stammte. Des weiteren wurde Plutonium in Urinproben bestimmt. Die Nachweisgrenzen lagen bei diesen Umweltproben bei 10^6 bis 10^7 Atomen Plutonium und damit um zwei Größenordnungen niedriger als die Nachweisgrenze für Pu-239 bei der alpha-Spektroskopie, der Standardmethode für den Plutoniumnachweis.

Entwicklung von Ionenquellen zur Optimierung von Selektivität und Effizienz bei der resonanten Laserionisation

In der vorliegenden Arbeit werden Entwicklungen zur Optimierung von Selektivität und Effizienz von Resonanzionisations-Laserionenquellen vorgestellt. Mit der Perspektive auf die Anwendungen radioaktiver Ionenstrahlen in der Grundlagenforschung sowie auf Fragestellungen in der Ultraspurenanalytik wurden verschiedene Methoden entwickelt und erprobt: Auf Seiten der Grundlagenforschung wurden zwei komplementäre Ansätze, die Konstruktion von Ionenquellen aus Materialien niedriger Austrittsarbeit und die Weiterentwicklung der Laserionenquelle und -falle LIST umgesetzt. Hierdurch konnte die Selektivität der Resonanzionisation in on-line Tests um einige Gröÿenordnungen verbessert werden. Für die Ultraspurenanalytik wurden speziell angepasste, hocheffiziente Ionenquellen entwickelt. Mit diesen Ionenquellen wurde für die Resonanzionisation von Gallium eine Ionisationseffizienz von 67 % demonstriert, für den Ultraspurennachweis des im Zusammenhang der nuklearen Endlagerung wichtigen Radioisotops 99g-Technetium wurde auf dieser Grundlage eine Nachweisgrenze von weniger als 10^6 Atomen gezeigt.

Implementierung der Laserionenquellenfalle LIST bei ISOLDE und Validierung der Spezifikationen Effizienz und Selektivität

Die Funktionsweise der Laserionenquellenfalle LIST, sowie deren Implementierung bei der Forschungseinrichtung ISOLDE am CERN als neue Standard-Ionenquelle und die ermittelten Spezifikationen Effizienz und Selektivität werden vorgestellt.rnrnDurch die Implementierung der LIST bei ISOLDE konnte on-line mit Hilfe von Radionukliden ein Minimalwert zur Unterdrückung von Kontaminationen durch die LIST bestimmt werden. Die erfolgreiche Unterdrückung von Francium-Kontamination ermöglichte es, neue Messdaten für den mittleren Ladungsradius und die Hyperfeinstruktur für Po-217 zu erzeugen.rnrnUm die Funktionalität der LIST bei ISOLDE hinsichtlich der Ionisationseffizienz gegenüber anderen Ionenquellen einzuordnen, wurden in Mainz am RISIKO-Massenseparator mit der bereits existierenden Standard-Ionenquelle RILIS und der LIST die Effizienzen bestimmt und miteinander verglichen. Es konnte gezeigt werden, dass die LIST im Modus hoher Ionisationseffizienz eine vergleichbare Effizienz aufweist wie die RILIS. Im Modus zur Produktion eines hochreinen Ionenstrahls ist die Ionisationseffizienz gegenüber der RILIS reduziert.rnrnDa die Bestimmung der Selektivität im On-line-Betrieb aufwendig und zeitintensiv ist, wurde die Reinheit des Ionenstrahls am RISIKO-Massenseparator mittels Laufzeitmessungen der Ionen off-line bestimmt und analysiert. Die Zeitstrukturen der RILIS ermöglichen einerseits Rückschlüsse auf die Reinheit des Ionenstrahls zu ziehen, andererseits konnte auch die Ionisation außerhalb des Atomisators, in dem überwiegend die resonante Ionisation stattfindet, nachgewiesen werden. Durch diesen Nachweis kann der Effizienzverlust während der Produktion eines hochreinen Ionenstrahls erklärt werden. Zudem bietet er einen Ansatz für weitere Entwicklungsarbeiten der LIST zur Steigerung der Effizienz.rnrnEine Übertragung der Messergebnisse zur Zeitstruktur der RILIS auf die LIST eröffnet neue Möglichkeiten zur Steigerung deren Selektivität im massenselektiven Mode. Dieser wurde anhand von Simulationen überprüft und mit Messungen an Kalium experimentell quantifiziert.

Extrapolated difference technique for the determination of atomization energies of Sm, Eu, and Yb bromides

An approach for the determination of atomization energies based on the extrapolated difference technique in the framework of Knudsen effusion mass spectrometry is proposed. Its essence is the use of thermodynamic data for the determination of the appearance energy of fragment ions of a reference and a special mathematical treatment of the ionization efficiency functions. The advantages of this approach are demonstrated for the cases of incongruently vaporizing lanthanide bromides that suffer from decomposition or disproportionation at high temperatures. The atomization energies for SmBr2 (7.78±0.12 eV), EuBr2 (7.51±0.11 eV), YbBr2 (7.25±0.13 eV), SmBr3 (11.09±0.10 eV), and YbBr3 (10.23±0.09 eV) molecules have been determined for the first time.

Knudsen effusion mass spectrometric determination of the complex vapor composition of samarium, europium, and ytterbium bromides

RATIONALE The vaporization of Sm, Eu, and Yb tri- and dibromides is accompanied by decomposition and disproportionation reactions. These result in complex vapor compositions whose analysis is an intricate problem for experimentalists. Approaches have been developed to interpret mass spectra and accurately determine the vapor composition of thermally unstable compounds. METHODS A sector type magnet instrument was used. A combined ion source allowed the study of both the molecular and ionic vapor compositions in the electron ionization (EI) and the thermionic emission (TE) modes. The methodological approaches were based on a joint analysis of the ionization efficiency functions, the temperature and time dependences of the ion currents, and special mathematical data evaluation. RESULTS The vaporization of SmBr3, YbBr3, SmBr2, EuBr2, and YbBr2 was studied in the temperature range of 850–1300 K. An initial stage of incongruent vaporization was observed in the case of the tribromides, SmBr2, and YbBr2. This eventually changed to a congruent vaporization stage. Various neutral (Ln, Br, Br2, LnBr, LnBr2, LnBr3, Ln2Br4, Ln2Br5, and Ln2Br6) and charged (Br–, LnBr3–, LnBr4–) species were detected at different vaporization stages. CONCLUSIONS The quantitative vapor composition of Sm, Eu, and Yb tri- and dibromides was determined. It was found that only EuBr2 was stable in the studied temperature range. The developed approaches can be useful in the case of other thermally unstable compounds.

Vapor phase composition as an indicator of thermal stability of the Sm, Eu, and Yb chlorides

The Sm, Eu, and Yb tri- and dichlorides were investigated by Knudsen effusion mass spectrometry. It was found out by the analysis of mass spectra and ionization efficiency curves that the vapor composition is complex due to the partial high temperature decomposition/disproportionation of the samples. Up to five vapor species were identified for both LnCl3 (LnCl3, LnCl2, Ln2Cl4, Ln2Cl5, and Ln2Cl6) and LnCl2 (LnCl3, LnCl2, LnCl, Ln, and Ln2Cl4). The quantitative evaluation of vapor composition was made. It indicates that the disproportionation of SmCl2 and EuCl2 is negligible in the temperature range studied whereas that of YbCl2 and the decomposition of SmCl3 and YbCl3 cannot be neglected.

Characterization of the Axial Jet Separator with a CO2/Helium Mixture: Toward GC-AMS Hyphenation

Development of interfaces for sample introduction from high pressures is important for real-time online hyphenation of chromatographic and other separation devices with mass spectrometry (MS) or accelerator mass spectrometry (AMS). Momentum separators can reduce unwanted low-density gases and introduce the analyte into the vacuum. In this work, the axial jet separator, a new momentum interface, is characterized by theory and empirical optimization. The mathematical model describes the different axial penetration of the components of a jetgas mixture and explains the empirical results for injections of CO2 in helium into MS and AMS instruments. We show that the performance of the new interface is sensitive to the nozzle size, showing good qualitative agreement with the mathematical model. Smaller nozzle sizes are more preferable due to their higher inflow capacity. The CO2 transmission efficiency of the interface into a MS instrument is ~14% (CO2/helium separation factor of 2.7). The interface receives and delivers flows of ~17.5 mL/min and ~0.9 mL/min, respectively. For the interfaced AMS instrument, the ionization and overall efficiencies are 0.7-3% and 0.1-0.4%, respectively, for CO2 amounts of 4-0.6 µg C, which is only slightly lower compared to conventional systems using intermediate trapping. The ionization efficiency depends on to the carbon mass flow in the injected pulse and is suppressed at high CO2 flows. Relative to a conventional jet separator, the transmission efficiency of the axial jet separator is lower, but its performance is less sensitive to misalignments.

(Table 3) Li concentrations and isotopic compositions in foraminiferal shells, carbonate sediments, and associated pore waters from ODP Sites 131-806 and 138-851

An improved procedure for lithium isotope analysis using Li3PO4 as the ion source has been investigated for application to geological samples. The 7Li/6Li ratio is measured using double rhenium filament thermal ionization mass spectrometry in which isotopic fractionation is minimized at high temperatures. The method produces a stable, high intensity Li+ ion beam that allows measurement of nanogram quantities of lithium. This results in a reduction in sample size of up to 1000 times relative to that required for the established Li2BO2+ method while maintaining a comparable precision of better than 1? (1 sigma). Replicate analyses of the NBS L-SVEC Li2CO3 standard yielded a mean value of 12.1047+/-0.0043 (n=21), which is close to the reported absolute value of 12.02+/-0.03. Intercalibration with a wide range of geological samples shows excellent agreement between the Li3PO4 and Li2BO2+ techniques. Replicate analyses of seawater and a fresh submarine basalt display high precision results that agree with previous measurements. Taking advantage of the high ionization efficiency of the phosphate ion source, we have made the first measurements of the lithium concentration (by isotope dilution) and isotopic composition of calcareous foraminiferal tests and other marine carbonates. Preliminary results indicate that substantial lithium exchange occurs between carbonate sediments and their interstitial waters. In addition, a possible link between lithium paleoceanography and paleoclimate during the last 1000 ky may be derived from planktonic foraminiferal tests. This highly sensitive technique can be applied in the examination of low lithium reservoirs and thereby provide insight into some fundamental aspects of lithium geochemistry.

Matrix effects: The Achilles heel of quantitative high-performance liquid chromatography-electrospray-tandem mass spectrometry

High-performance liquid chromatography coupled by an electrospray ion source to a tandem mass spectrometer (HPLC-EST-MS/ MS) is the current analytical method of choice for quantitation of analytes in biological matrices. With HPLC-ESI-MS/MS having the characteristics of high selectivity, sensitivity, and throughput, this technology is being increasingly used in the clinical laboratory. An important issue to be addressed in method development, validation, and routine use of HPLC-ESI-MS/MS is matrix effects. Matrix effects are the alteration of ionization efficiency by the presence of coeluting substances. These effects are unseen in the chromatograrn but have deleterious impact on methods accuracy and sensitivity. The two common ways to assess matrix effects are either by the postextraction addition method or the postcolumn infusion method. To remove or minimize matrix effects, modification to the sample extraction methodology and improved chromatographic separation must be performed. These two parameters are linked together and form the basis of developing a successful and robust quantitative HPLC-EST-MS/MS method. Due to the heterogenous nature of the population being studied, the variability of a method must be assessed in samples taken from a variety of subjects. In this paper, the major aspects of matrix effects are discussed with an approach to address matrix effects during method validation proposed. (c) 2004 The Canadian Society of Clinical Chemists. All rights reserved.

SU-8-Based Microchips for Capillary Electrophoresis and Electrospray Ionization Mass Spectrometry

Miniaturization of analytical instrumentation is attracting growing interest in response to the explosive demand for rapid, yet sensitive analytical methods and low-cost, highly automated instruments for pharmaceutical and bioanalyses and environmental monitoring. Microfabrication technology in particular, has enabled fabrication of low-cost microdevices with a high degree of integrated functions, such as sample preparation, chemical reaction, separation, and detection, on a single microchip. These miniaturized total chemical analysis systems (microTAS or lab-on-a-chip) can also be arrayed for parallel analyses in order to accelerate the sample throughput. Other motivations include reduced sample consumption and waste production as well as increased speed of analysis. One of the most promising hyphenated techniques in analytical chemistry is the combination of a microfluidic separation chip and mass spectrometer (MS). In this work, the emerging polymer microfabrication techniques, ultraviolet lithography in particular, were exploited to develop a capillary electrophoresis (CE) separation chip which incorporates a monolithically integrated electrospray ionization (ESI) emitter for efficient coupling with MS. An epoxy photoresist SU-8 was adopted as structural material and characterized with respect to its physicochemical properties relevant to chip-based CE and ESI/MS, namely surface charge, surface interactions, heat transfer, and solvent compatibility. As a result, SU-8 was found to be a favorable material to substitute for the more commonly used glass and silicon in microfluidic applications. In addition, an infrared (IR) thermography was introduced as direct, non-intrusive method to examine the heat transfer and thermal gradients during microchip-CE. The IR data was validated through numerical modeling. The analytical performance of SU-8-based microchips was established for qualitative and quantitative CE-ESI/MS analysis of small drug compounds, peptides, and proteins. The CE separation efficiency was found to be similar to that of commercial glass microchips and conventional CE systems. Typical analysis times were only 30-90 s per sample indicating feasibility for high-throughput analysis. Moreover, a mass detection limit at the low-attomole level, as low as 10E+5 molecules, was achieved utilizing MS detection. The SU-8 microchips developed in this work could also be mass produced at low cost and with nearly identical performance from chip to chip. Until this work, the attempts to combine CE separation with ESI in a chip-based system, amenable to batch fabrication and capable of high, reproducible analytical performance, have not been successful. Thus, the CE-ESI chip developed in this work is a substantial step toward lab-on-a-chip technology.

Miniaturized mass spectrometric ionization techniques for environmental analysis and bioanalysis

Miniaturized mass spectrometric ionization techniques for environmental analysis and bioanalysis Novel miniaturized mass spectrometric ionization techniques based on atmospheric pressure chemical ionization (APCI) and atmospheric pressure photoionization (APPI) were studied and evaluated in the analysis of environmental samples and biosamples. The three analytical systems investigated here were gas chromatography-microchip atmospheric pressure chemical ionization-mass spectrometry (GC-µAPCI-MS) and gas chromatography-microchip atmospheric pressure photoionization-mass spectrometry (GC-µAPPI-MS), where sample pretreatment and chromatographic separation precede ionization, and desorption atmospheric pressure photoionization-mass spectrometry (DAPPI-MS), where the samples are analyzed either as such or after minimal pretreatment. The gas chromatography-microchip atmospheric pressure ionization-mass spectrometry (GC-µAPI-MS) instrumentations were used in the analysis of polychlorinated biphenyls (PCBs) in negative ion mode and 2-quinolinone-derived selective androgen receptor modulators (SARMs) in positive ion mode. The analytical characteristics (i.e., limits of detection, linear ranges, and repeatabilities) of the methods were evaluated with PCB standards and SARMs in urine. All methods showed good analytical characteristics and potential for quantitative environmental analysis or bioanalysis. Desorption and ionization mechanisms in DAPPI were studied. Desorption was found to be a thermal process, with the efficiency strongly depending on thermal conductivity of the sampling surface. Probably the size and polarity of the analyte also play a role. In positive ion mode, the ionization is dependent on the ionization energy and proton affinity of the analyte and the spray solvent, while in negative ion mode the ionization mechanism is determined by the electron affinity and gas-phase acidity of the analyte and the spray solvent. DAPPI-MS was tested in the fast screening analysis of environmental, food, and forensic samples, and the results demonstrated the feasibility of DAPPI-MS for rapid screening analysis of authentic samples.

Development of atmospheric pressure ionization ion mobility spectrometry and ion mobility spectrometry mass spectrometry

This study is focused on the development and evaluation of ion mobility instrumentation with various atmospheric pressure ionization techniques and includes the following work. First, a high-resolution drift tube ion mobility spectrometer (IMS), coupled with a commercial triple quadrupole mass spectrometer (MS), was developed. This drift tube IMS is compatible with the front-end of commercial Sciex mass spectrometers (e.g., Sciex API-300, 365, and 3000) and also allows easy (only minor modifications are needed) installation between the original atmospheric pressure ion source and the triple quadrupole mass spectrometer. Performance haracteristics (e.g.,resolving power, detection limit, transmission efficiency of ions) of this IMS-MS instrument were evaluated. Development of the IMS-MS instrument also led to a study where a proposal was made that tetraalkylammonium ions can be used as chemical standards for ESI-IMS. Second, the same drift tube design was also used to build a standalone ion mobility spectrometer equipped with a Faraday plate detector. For this highresolution (resolving power about 100 shown) IMS device, a multi-ion source platform was built, which allows the use of a range of atmospheric pressure ionization methods, such as: corona discharge chemical ionization (CD-APCI), atmospheric pressure photoionization (APPI), and radioactive atmospheric pressure chemical ionization (R-APCI). The multi-ion source platform provides easy switching between ionization methods and both positive and negative ionization modes can be used. Third, a simple desorpion/ionization on silicon (DIOS) ion source set-up for use with the developed IMS and IMS-MS instruments was built and its operation demonstrated. Fourth, a prototype of a commercial aspiration-type ion mobility spectrometer was mounted in front of a commercial triple quadrupole mass spectrometer. The set-up, which is simple, easy to install, and requires no major modifications to the MS, provides the possibility of gathering fundamental information about aspiration mobility spectrometry.