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A mobile mass spectrometer for comprehensive on-line analysis of trace and bulk components of complex gas mixtures: parallel application of the laser-based ionization methods VUV single-photon ionization, resonant multiphoton ionization, and laser-induced electron impact ionization.

A newly developed compact and mobile time-of-flight mass spectrometer (TOFMS) for on-line analysis and monitoring of complex gas mixtures is presented. The instrument is designed for a (quasi-)simultaneous application of three ionization techniques that exhibit different ionization selectivities. The highly selective resonance-enhanced multiphoton ionization (REMPI) technique, using 266-nm UV laser pulses, is applied for selective and fragmentationless ionization of aromatic compounds at trace levels (parts-per-billion volume range). Mass spectra obtained using this technique show the chemical signature solely of monocyclic (benzene, phenols, etc.) and polycyclic (naphthalene, phenathrene, indol, etc.) aromatic species. Furthermore, the less selective but still fragmentationless single photon ionization (SPI) technique with 118-nm VUV laser pulses allows the ionization of compounds with an ionization potential below 10.5 eV. Mass spectra obtained using this technique show the profile of most organic compounds (aliphatic and aromatic species, like nonane, acetaldehyde, or pyrrol) and some inorganic compounds (e.g., ammonia, nitrogen monoxide). Finally, the nonselective ionization technique laser-induced electron-impact ionization (LEI) is applied. However, the sensitivity of the LEI technique is adjusted to be fairly low. Thus, the LEI signal in the mass spectra gives information on the inorganic bulk constituents of the sample (i.e., compounds such as water, oxygen, nitrogen, and carbon dioxide). Because the three ionization methods (REMPI, SPI, LEI) exhibit largely different ionization selectivities, the isolated application of each method alone solely provides specific mass spectrometric information about the sample composition. Special techniques have been developed and applied which allow the quasi-parallel use of all three ionization techniques for on-line monitoring purposes. Thus, a comprehensive characterization of complex samples is feasible jointly using the characteristic advantages of the three ionization techniques. Laboratory applications show results on rapid overview characterization of mineral oil-based fuels and coffee headspace. The first reported field applications include timely resolved on-line monitoring results on automobile exhausts and of waste incineration flue gas.

Journal Article↗

Ionization enhancement in atmospheric pressure chemical ionization and suppression in electrospray ionization between target drugs and stable-isotope-labeled internal standards in quantitative liquid chromatography/tandem mass spectrometry.

The phenomena of ionization suppression in electrospray ionization (ESI) and enhancement in atmospheric pressure chemical ionization (APCI) were investigated in selected-ion monitoring and selected-reaction monitoring modes for nine drugs and their corresponding stable-isotope-labeled internal standards (IS). The results showed that all investigated target drugs and their co-eluting isotope-labeled IS suppress each other's ionization responses in ESI. The factors affecting the extent of suppression in ESI were investigated, including structures and concentrations of drugs, matrix effects, and flow rate. In contrast to the ESI results, APCI caused seven of the nine investigated target drugs and their co-eluting isotope-labeled IS to enhance each other's ionization responses. The mutual ionization suppression or enhancement between drugs and their isotope-labeled IS could possibly influence assay sensitivity, reproducibility, accuracy and linearity in quantitative liquid chromatography/mass spectrometry (LC/MS) and liquid chromatography/tandem mass spectrometry (LC/MS/MS). However, calibration curves were linear if an appropriate IS concentration was selected for a desired calibration range to keep the response factors constant.

Artifacts↗

Resonance-enhanced multiphoton ionization and VUV-single photon ionization as soft and selective laser ionization methods for on-line time-of-flight mass spectrometry: investigation of the pyrolysis of typical organic contaminants in the steel recycling process.

A newly conceived compact and mobile time-of flight mass spectrometer (TOFMS) for real-time monitoring of highly complex gas mixtures is presented. The device utilizes two selective and sensitive soft ionization techniques, viz., resonance-enhanced multiphoton ionization (REMPI) and single-photon ionization (SPI) in a (quasi)-simultaneous mode. Both methods allow a fragmentationless ionization. The REMPI method selectively addresses aromatic species, while with SPI applying vacuum ultaviolet light (118 nm) in principle all compounds with an ionization potential below 10.5 eV are accessible. This provides comprehensive information of the chemical composition of complex matrixes. The combustion and pyrolysis behavior of five organic materials typically used in steel processing in China was studied. The trace amounts of organic compounds in the gas phase during combustion and pyrolysis were monitored selectively and sensitively by real-time SPI/REMPI-TOFMS. The measurements were carried out at several constant temperatures in the range from 300 to 1190 degrees C in both synthetic air and nitrogen. Timely resolved mass spectra reveal the formation and subsequent growth of aromatic molecules. At lower temperatures, highly alkylated PAHs predominate, while at temperatures above 800 degrees C, the more stable benzene and PAHs without side chains prevail. Potential hyphenation of SPI/REMPI-TOFMS to methods of thermal analysis is discussed.

Journal Article↗

Identification of acetylation and methylation sites of histone H3 from chicken erythrocytes by high-accuracy matrix-assisted laser desorption ionization-time-of-flight, matrix-assisted laser desorption ionization-postsource decay, and nanoelectrospray ionization tandem mass spectrometry.

A new strategy has been employed for the identification of the covalent modification sites (mainly acetylation and methylation) of histone H3 from chicken erythrocytes using low enzyme/substrate ratios and short digestion times (trypsin used as the protease) with analysis by HPLC separation, matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF), matrix-assisted laser desorption ionization-postsource decay, and tandem mass spectrometric techniques. High-accuracy MALDI-TOF mass measurements with representative immonium ions (126 for acetylated lysine, 98 for monomethylated lysine, and 84 for di-, tri-, and unmethylated lysine) have been effectively used for differentiating methylated peptides from acetylated peptides. Our results demonstrate that lysines 4, 9, 14, 27, and 36 of the N-terminal of H3 are methylated, while lysines 14, 18, and 23 are acetylated. Surprisingly, a non-N-terminal residue, lysine 79, in the loop region hooking up to the bound DNA, was newly found to be methylated (un-, mono-, and dimethylated isoforms coexist). The reported mass spectrometric method has the advantages of speed, directness, sensitivity, and ease over protein sequencing and Western-blotting methods and holds the promise of an improved method for determining the status of histone modifications in the field of chromosome research.

Acetylation↗

Liquid chromatography/mass spectrometry in anabolic steroid analysis--optimization and comparison of three ionization techniques: electrospray ionization, atmospheric pressure chemical ionization and atmospheric pressure photoionization.

The applicability of liquid chromatography/tandem mass spectrometry (LC/MS/MS) for the detection of the free anabolic steroid fraction in human urine was examined. Electrospray ionization (ESI), atmospheric pressure chemical ionization and atmospheric pressure photoionization methods were optimized regarding eluent composition, ion source parameters and fragmentation. The methods were compared with respect to specificity and detection limit. Although all methods proved suitable, LC/ESI-MS/MS with a methanol-water gradient including 5 mM ammonium acetate and 0.01% acetic acid was found best for the purpose. Multiple reaction monitoring allowed the determination of steroids in urine at low nanogram per milliliter levels. LC/MS/MS exhibited high sensitivity and specificity for the detection of free steroids and may be a suitable technique for screening for the abuse of anabolic steroids in sports.

Anabolic Agents↗

Solubilization of ionized and un-ionized flavopiridol by ethanol and polysorbate 20.

Because the ionized species is more polar than its un-ionized counterpart, it is often assumed that the ionized species of the drug does not make a meaningful contribution to solubilization by either cosolvents or surfactants. This report extends previous studies on solubilization of the ionic species by a combination of pH control and complexation to pH control and micellization and to pH control and cosolvency. The total aqueous solubility is expressed as the addition of the concentration of all contributing species: free un-ionized drug [Du], free ionized drug [Di], un-ionized drug micelle [DuM], and ionized drug micelle [DiM] for surfactant, and free un-ionized drug [Dcu] and free ionized drug [Dci] for cosolvent. The equations indicate that under certain conditions the ionized species can be more important in determining the drug total solubility than the un-ionized species. Flavopiridol, a weak base, is used to test these newly generated equations. As expected, the micellar partition coefficient and solubilization power for ionized flavopiridol are both less than those of the un-ionized species. However, at acidic pH, the solubilities of the ionized drug in surfactant micelles [DiM] and in cosolvent-water [Dci] are both much greater than that of the un-ionized drug. This difference is because the solubilization of the ionized drug is proportional to its aqueous solubility, and its solubility [Di] can be as much as 24-fold greater than that of the free un-ionized species [Du].

Antineoplastic Agents↗

Whole blood ionized magnesium: age-related differences in normal values and clinical implications of ionized hypomagnesemia in patients undergoing surgery for congenital cardiac disease.

OBJECTIVES: We sought to (1) determine reference values for whole blood ionized magnesium concentrations in newborns, children, and young adults and (2) evaluate the frequency and clinical implications of ionized hypomagnesemia in patients undergoing surgery for congenital heart disease. METHOD: We prospectively measured ionized magnesium concentrations in 299 subjects (113 control subjects and 186 patients undergoing surgery for congenital heart disease). Subjects were categorized by age. In the surgical group blood samples were obtained before bypass, during bypass (cooling and rewarming), after bypass, and during admission to the intensive care unit. Ionized hypomagnesemia was defined as ionized magnesium level 2 standard deviations below the mean of control subjects in the same age group. Patients were analyzed, controlling for cardiopulmonary bypass time. RESULTS: In the control group ionized magnesium concentrations differed by age. Neonates and adults showed lower ionized magnesium concentrations compared with those of other age groups. Infants exhibited the highest ionized magnesium concentration. In the surgical group patients older than 1 month showed a higher proportion of ionized hypomagnesemia compared with that found in neonates at baseline (P <.001), after bypass (P =. 03), and at admission to the intensive care unit (P =.02). Controlling for cardiopulmonary bypass time, patients older than 1 month who were hypomagnesemic during bypass showed longer intubation time (P =.001) and longer intensive care stay (P =.01) and tended to have a higher pediatric severity of illness score on intensive care admission (P =.14) compared with patients without ionized hypomagnesemia. CONCLUSIONS: There are age-related differences in normal ionized magnesium concentrations. Ionized hypomagnesemia is a common and clinically relevant occurrence among patients older than 1 month of age undergoing surgery for congenital heart disease.

Adolescent↗

pH effects on measurements of ionized calcium and ionized magnesium in blood.

CONTEXT: It is well known that the concentration of ionized calcium in blood is affected by the pH of the specimen, since hydrogen ions compete with calcium for binding sites on albumin and other proteins. However, the relationship between pH and ionized magnesium concentration is not as well characterized. OBJECTIVE: To determine the effects of pH on ionized magnesium concentration over a wide range of pH values in serum or plasma. DESIGN: Both ionized calcium and ionized magnesium concentrations were measured in 3 sets of samples. (1) Pools of serum or whole blood at different pH values (7.20-7.60) achieved by adding a constant volume of acid or base (diluted solutions of either hydrochloric acid or sodium hydroxide) plus saline. These pools consisted of 2 serum and 3 heparinized whole blood pools collected from leftover blood remaining in clinical specimens in the Clinical Chemistry and Blood Gas Laboratories, respectively, at Duke University Medical Center. (2) Five whole blood specimens obtained from apparently healthy individual donors. (3) Twenty-six whole blood specimens obtained from individual patients (leftover blood from the Blood Gas Laboratory) in which pH was varied by in vitro loss or gain of carbon dioxide. RESULTS: Both ionized calcium and ionized magnesium concentrations decreased as the pH in the specimen increased, indicating the stronger binding of these ions with proteins in the more alkaline environment. CONCLUSION: We conclude that the rate of change of ionized magnesium concentration with pH change (0.12 mmol/L per pH unit) is significantly less than that of ionized calcium (0.36 mmol/L per pH unit). Furthermore, our findings indicate that if adjustment to pH 7.40 is necessary, the ionized magnesium test results need to be adjusted when pH is markedly abnormal, as is sometimes done for ionized calcium.

Calcium↗

Ionized magnesium supplementation in critically ill patients: comparing ionized and total magnesium.

PURPOSE: The purpose of this study was to assess the effect of magnesium supplementation on total magnesium, ionized magnesium, ionized calcium, potassium, and pH in critically ill cancer patients and to compare the validity of the measurements. MATERIALS AND METHODS: Thirty-three consecutive critically ill patients receiving magnesium supplementation were placed in this prospective observational study at the Comprehensive Cancer Center, University Hospital. One gram (4.1 mmol) magnesium in 50 mL D5W was administered to critically ill patients, and the following were measured: total magnesium, ionized magnesium, ionized calcium, potassium, albumin, pH, BUN, creatinine, creatinine. RESULTS: Total magnesium and ionized magnesium increased by a mean of .11 +/- .02 and .05 +/- .01 mmol/L, respectively, after supplementation with 4.1 mmol of magnesium sulfate (P = .0001). Total magnesium, ionized magnesium, albumin, ionized calcium, potassium, and pH did not change significantly by the administration of 1 g of magnesium sulfate. CONCLUSION: The mean ionized magnesium (IMg+2) relationship to total magnesium (TMg) cannot be predicted before the supplementation with the available technology. After supplementation of 4.1 mmol/L the ionized magnesium level increased by .05 +/- .01 mmol/L. Magnesium supplementation had no significant effect on ionized calcium, potassium, and pH. TMg and IMg+2 should be followed independently.

Adult↗

Atmospheric pressure photoionization mass spectrometry. Ionization mechanism and the effect of solvent on the ionization of naphthalenes.

The ionization mechanism in dopant-assisted atmospheric pressure photoionization and the effect of solvent on the ionization efficiency was studied using 7 naphthalenes and 13 different solvent systems. The ionization efficiency was 1-2 orders of magnitude higher with dopant than without, indicating that the photoionization of the dopant initiates the ionization process. In positive ion mode, the analytes were ionized either by charge exchange or by proton transfer. Charge exchange was favored for low proton affinity solvents (water, hexane, chloroform), whereas the addition of methanol or acetonitrile to the solvent initiated proton transfer. In negative ion mode, the compounds with high electron affinity were ionized by electron capture or by charge exchange and the compounds with high gas-phase acidity were ionized by proton transfer. In addition, some oxidation reactions were observed. All the reactions leading to ionization of analytes in negative ion mode are initiated by thermal electrons formed in photoionization of toluene. The testing of different solvents showed that addition of buffers such as ammonium acetate, ammonium hydroxide, or acetic acid may suppress ionization in APPI. The reactions are discussed in detail in light of thermodynamic data.

Atmospheric Pressure↗

The simultaneous measurement of ionized and total calcium and ionized and total magnesium in intensive care unit patients.

PURPOSE: This study was undertaken to determine the relationship between total magnesium and ionized magnesium in critically ill and injured patients. METHODS: Eighty consecutive intensive care unit (ICU) admissions were evaluated and 34 patients were enrolled in the study. Patients were enrolled who had indwelling arterial catheters and were within 4 days of ICU admission. Six milliliters of blood was collected and assayed simultaneously for total and ionized magnesium, total and ionized calcium, and albumin level. An Acute Physiology and Chronic Health Evaluation (APACHE II) score was calculated at the time of blood collection. RESULTS: The results of our study show a strong correlation between ionized and total magnesium (R =.903) that was not seen between ionized and total calcium (R =.748). We found total hypomagnesemia in 18% and ionized hypomagnesemia in 21% of ICU patients. We also found that 14.7% (5 of 34) of our patients had ionized hypermagnesemia whereas none displayed total hypermagnesemia. We did not find a correlation between APACHE II, sex, race, albumin level, and any electrolyte level. The mortality rate in the subjects studied was 21% (7 of 34). CONCLUSIONS: Based on our results we would recommend that intensivists directly measure ionized calcium whereas ionized magnesium can be inferred from total magnesium.

APACHE↗

Specific heparin preparations interfere with the simultaneous measurement of ionized magnesium and ionized calcium.

OBJECTIVE: To determine whether heparin anticoagulants used for analysis of whole blood ionized calcium would influence the measurement of ionized magnesium. METHODS: The effects of zinc heparin, lithium heparin, and electrolyte-balanced heparin on the simultaneous measurement of ionized magnesium and ionized calcium in serum were determined using ion selective electrodes. RESULTS: Time-dependent biases in ionized magnesium and calcium concentrations were apparent with zinc heparin but not with lithium or electrolyte-balanced heparin. Ionized magnesium and calcium concentrations were more significantly influenced by volume-dependent changes in zinc heparin potency than with lithium or electrolyte-balanced heparin. CONCLUSION: Zinc heparin produces a significant positive bias in the simultaneous determination of ionized magnesium and ionized calcium concentrations.

Anticoagulants↗

Atmospheric pressure chemical ionization studies of non-polar isomeric hydrocarbons using ion mobility spectrometry and mass spectrometry with different ionization techniques.

The ionization pathways were determined for sets of isomeric non-polar hydrocarbons (structural isomers, cis/trans isomers) using ion mobility spectrometry and mass spectrometry with different techniques of atmospheric pressure chemical ionization to assess the influence of structural features on ion formation. Depending on the structural features, different ions were observed using mass spectrometry. Unsaturated hydrocarbons formed mostly [M - 1]+ and [(M - 1)2H]+ ions while mainly [M - 3]+ and [(M - 3)H2O]+ ions were found for saturated cis/trans isomers using photoionization and 63Ni ionization. These ionization methods and corona discharge ionization were used for ion mobility measurements of these compounds. Different ions were detected for compounds with different structural features. 63Ni ionization and photoionization provide comparable ions for every set of isomers. The product ions formed can be clearly attributed to the structures identified. However, differences in relative abundance of product ions were found. Although corona discharge ionization permits the most sensitive detection of non-polar hydrocarbons, the spectra detected are complex and differ from those obtained with 63Ni ionization and photoionization.

Atmospheric Pressure↗

A two-color three-photon ionization scheme for the efficient and selective ionization of a chlorinated aromatic hydrocarbon.

A two-color three-photon ionization scheme, for the efficient and selective ionization of a chlorinated aromatic hydrocarbon that has an ionization potential higher than the two-photon energy of the laser used for excitation, is described. In this technique, an ultraviolet (UV) laser, i.e., the second harmonic emission of a fundamental (VIS) laser, is used for excitation and a UV and VIS laser for the subsequent two-photon ionization from the electronic excited state. A sample of o-chlorophenol was used as a model compound to demonstrate the advantage of this technique. The signal in supersonic jet/resonance-enhanced multiphoton ionization/mass spectrometry was increased approximately 4 times by the introduction of the VIS beam, when the polarization was adjusted to be parallel to the UV beam. Thus, the two-color three-photon (2UV+VIS) ionization scheme is more sensitive than one-color three-photon (3UV) ionization. The merits of this method over other ionization schemes such as two-color two-photon (UV(1)+UV(2)) ionization are discussed in terms of sensitivity and selectivity in spectrometric analysis.

Letter↗

Adjusted ionized calcium (at pH 7.4) and actual ionized calcium (at actual pH) in capillary blood compared for clinical evaluation of patients with disorders of calcium metabolism.

We report results for adjusted ionized calcium (at pH 7.4) and actual ionized calcium (at actual pH) in capillary blood from 183 patients with disorders of calcium metabolism (primary hyperparathyroidism, secondary hyperparathyroidism of malabsorption, primary hypoparathyroidism, Paget's disease, acromegaly, hypercalcemia of malignancy, osteoporosis, sarcoidosis, idiopathic hypercalciuria, and familial hypocalciuric hypercalcemia). The correlation and the equation for the linear regression between adjusted ionized calcium (y) and actual ionized calcium (x) were y = 1.011x + 0.005 mmol/L, r = 0.992, Sy,x = 0.021 mmol/L. Results were similar within each diagnostic group. Consistent agreement between adjusted and ionized calcium was observed in 96.7% of patients representing a variety of the most frequently encountered disorders of calcium metabolism. Thus we find adjusted ionized calcium to be as useful as actual ionized calcium for evaluation of patients with such disorders. Adjusted ionized calcium may therefore also be a logical choice for establishing agreement between laboratories for reference intervals in healthy adults.

Acromegaly↗

Influence of the eluent composition on the ionization efficiency for morphine of pneumatically assisted electrospray, atmospheric-pressure chemical ionization and sonic spray.

A comparative study of three atmospheric-pressure ionization (API) sources for liquid chromatography/mass spectrometry (LC/MS), namely pneumatically assisted electrospray or ionspray (IS), atmospheric-pressure chemical ionization (APCI), and sonic spray (SS), with respect to the influence of the eluent composition on the ionization of morphine, is presented. The effect of organic modifiers, volatile acids, and buffer systems (with and without pH adjustment) in the LC mobile phase on the ionization efficiency of each interface is described. We conclude that for all three ion sources, the composition of the liquid phase had a serious impact on the ionization of the target compound. For IS and SS, very similar behavior towards the LC eluent was observed. In both cases, an increase in organic modifier resulted in an increase in ionization, while an increasing amount of volatile acid or buffer caused signal suppression. APCI, on the other hand, proved to respond completely differently towards the changes in the eluent. Again, an increased ionization was observed with an increase in organic modifier content but this time also in the presence of mobile phase additives like acids or buffers. Finally, we concluded that APCI proved to be the preferred ion source for the test compound because of its robust character and its direct applicability in traditional LC analysis.

Chromatography, Liquid↗

Chromatographic and ionization properties of polybrominated diphenyl ethers using GC/high-resolution MS with metastable atom bombardment and electron impact ionization.

The chromatographic and ionization properties of 35 polybrominated diphenyl ether (PBDE) congeners were investigated using GC/HRMS with metastable atom bombardment (MAB) and electron impact (EI) ionization. A multiple linear regression model based on bromine substitution patterns and MOPAC calculated physical properties was developed to predict relative GC retention times of individual PBDE congeners. Although five different sources of metastable rare gas atoms (He, N2, Ar, Xe, and Kr) were investigated with MAB ionization, only MAB-N2 provided adequate ionization efficiency and predictability. Because of reduced background noise to the MS detector, MAB-N2 had a lower limit of detection for tetra- and penta-BDEs than EI, despite having a lower sensitivity. Using MAB-N2, the molecular ion was always the base peak, with little fragmentation taking place. Conversely, using EI ionization, the [M - nBr]+ peak (where n = 1-4, depending on the number of Br substituents) was the dominant ion for all PBDE congeners. Multiple linear regression models representing the molecular ion response of PBDE congeners analyzed by GC/ HRMS with MAB-N2 and EI ionization were also developed using the number and type of Br substituents and ionization potentials. A significantly higher level of predictability was obtained for the MAB-N2 response model than for EI.

Electrons↗

Ionization energies of hypervalent Li2F, Li2Cl and Na2Cl molecules obtained by surface ionization electron impact neutralization mass spectrometry.

Ionization energies of hypervalent Li(2)F, Li(2)Cl and Na(2)Cl molecules detected by surface ionization electron impact neutralization mass spectrometry are reported. The ionization energies were 3.78 +/- 0.2 eV for Li(2)F, 4.93 +/- 0.2 eV for Li(2)Cl, and 4.21 +/- 0.2 eV for Na(2)Cl. The ionization energies (IE) agree with theoretical ionization energies calculated by ab initio methods, supporting the theoretical prediction that Li(2)F has a hyperlithiated configuration in which the odd electron delocalizes over the two lithiums and with photoionization measurement. The first ionization energy of Na(2)Cl was experimentally confirmed earlier and for Li(2)Cl as well.8 We have developed and used this new approach for the problem--in the present work ions were first formed by surface ionization, followed by electron attachment (neutralization).

Journal Article↗