Christopher R. Bowerbank

Determination of catecholamines and metanephrines in urine by capillary electrophoresis–electrospray ionization–time-of-flight mass spectrometry

A method successfully coupling capillary electrophoretic separation to time-of-flight mass spectrometric (TOFMS) detection for the simultaneous analysis of catecholamines (dopamine, norepinephrine, and epinephrine) and their O-methoxylated metabolites (3-methoxytyramine, normetanephrine, and metanephrine) is described. The inner capillary wall was coated with polyvinyl alcohol in order to obtain baseline resolution of catecholamines and metanephrines and to ensure reproducibility without extensive restorative washing of the capillary. Using electrokinetic injection, detection limits of 0.3 microM for dopamine and norepinephrine, 0.2 microM for 3-methoxytyramine and normetanephrine, and 0.1 microM for epinephrine and metanephrine were achieved with standard solutions. The usefulness of this approach was demonstrated by applying the developed method to the analysis of a spot collection of human urine from a healthy volunteer. The catecholamines and metanephrines were removed from the urine samples and preconcentrated by simultaneous SPE on cation-exchange sorbents. The recoveries of all analytes, with the exception of epinephrine (75%), were over 80%. Catecholamines and metanephrines in the urine samples were quantitated using 3,4-dihydroxybenzylamine as an internal standard. Submicromolar concentrations, consistent with the catecholamine and metanephrine levels reported for normal human urine, were detected.

Advantages and limitations of coupling isotachophoresis and comprehensive isotachophoresis-capillary electrophoresis to time-of-flight mass spectrometry.

Capillary isotachophoresis (ITP) and comprehensive isotachophoresis-capillary electrophoresis (ITP-CE) were successfully coupled to electrospray ionization (ESI) orthogonal acceleration time-of-flight mass spectrometry (TOF-MS) using angiotensin peptides as model analytes. The utility of ITP-TOF-MS and ITP-CE-TOF-MS for the analysis of samples containing analyte amounts sufficient to form flat-top ITP zones (30 microM) as well as for samples with trace analyte amounts (0.3 microM) was studied. Separations were performed in 150 microm internal diameter (I.D.) capillaries for the ITP experiments, and in 200 microm I.D. (ITP) and 50 microm I.D. (CE) capillaries for ITP-CE experiments. The fused-silica columns were coated with poly(vinyl alcohol) to suppress electroosmotic flow that can disrupt ITP zone profiles. The sample loading capacity in both ITP and comprehensive ITP-CE was greatly enhanced (up to 10 microl) compared with typical nanoliter-sized injection volumes in CE. It was concluded that ITP-TOF-MS alone was adequate for the separation and detection of high concentration samples. The outcome was different at lower analyte concentrations where mixed zones or very sharp peaks formed. With formation of mixed zones, ion suppression and discrimination could occur, complicating quantitative determination of the analytes. This problem was effectively overcome by inserting a CE capillary between the ITP and TOF-MS. In such an arrangement, samples were preconcentrated in the high load WTP capillary and then injected into a CE capillary where they were separated into non-overlapping peaks prior to their detection by TOF-MS. The advantage of this comprehensive arrangement, which we have described previously, is that there is no need to discard portions of the sample in order to avoid overloading of the CE capillary. The whole sample is analyzed by multiple injections from ITP to CE. Thus, this method can be used for the analysis of complex samples with wide ranges of component concentrations.

Use of a hand-portable gas chromatograph-toroidal ion trap mass spectrometer for self-chemical ionization identification of degradation products related to O-ethyl S-(2-diisopropylaminoethyl) methyl phosphonothiolate (VX)

The chemical warfare agent O-ethyl S-(2-diisopropylaminoethyl) methyl phosphonothiolate (VX) and many related degradation products produce poorly diagnostic electron ionization (EI) mass spectra by transmission quadrupole mass spectrometry. Thus, chemical ionization (CI) is often used for these analytes. In this work, pseudomolecular ([M+H](+)) ion formation from self-chemical ionization (self-CI) was examined for four VX degradation products containing the diisopropylamine functional group. A person-portable toroidal ion trap mass spectrometer with a gas chromatographic inlet was used with EI, and both fixed-duration and feedback-controlled ionization time. With feedback-controlled ionization, ion cooling (reaction) times and ion formation target values were varied. Evidence for protonation of analytes was observed under all conditions, except for the largest analyte, bis(diisopropylaminoethyl)disulfide which yielded [M+H](+) ions only with increased fixed ionization or ion cooling times. Analysis of triethylamine-d(15) provided evidence that [M+H](+) production was likely due to self-CI. Analysis of a degraded VX sample where lengthened ion storage and feedback-controlled ionization time were used resulted in detection of [M+H](+) ions for VX and several relevant degradation products. Dimer ions were also observed for two phosphonate compounds detected in this sample.

Solvating gas chromatography with chemiluminescence detection of nitroglycerine and other explosives

A separation technique known as solvating gas chromatography (SGC), which utilizes packed capillary columns and neat carbon dioxide as mobile phase, was used for the separation of nitroglycerine (NG) and other nitrogen-containing explosives including 2,6-dinitrotoluene (2,6-DNT), 2,4-dinitrotolulene (2,4-DNT), 2,4,6-trinitrotoluene (2,4,6-TNT), and pentaerythritol tetranitrate (PETN). SGC was coupled for the first time to a selective chemiluminescence thermal energy analyzer (TEA) detector for nitro-functional group specificity and sensitive detection of these compounds. TEA calibration curve for NG showed linearity in the sub-microg ml(-1) range. Soil samples containing NG were used to test the validity of the technique. Detector response of SGC-TEA versus SGC-flame ionization detection for NG was also evaluated.

Rapid separation of nitroaromatic compounds by solvating gas chromatography.

Nitroaromatic compounds such as 2,4,6-trinitrotoluene are used in the production of explosives and munitions, and are environmental contaminants as a result of such use. Conventional methods to detect these compounds have relied on liquid and gas chromatography to isolate the individual compounds which may be present at low levels in complex environmental matrices before final detection and identification. A new method, solvating gas chromatography, was used to rapidly separate 8 nitroaromatic compounds, showing improved speed relative to conventional liquid and gas chromatography methods. Solvating gas chromatography allows near real-time detection for these energetic compounds, providing improvements in their detection as environmental contaminants, and as compounds of interest to law enforcement and military organizations.

Airborne aldehydes from heating rosin core solder and liquid rosin flux to soldering temperatures.

Gas phase aldehydes produced from heating rosin core solder and liquid rosin flux to temperatures commonly used in soldering were trapped on sampling tubes containing XAD-2 resin coated with the derivatizing agent 2-hydroxymethylpiperidine. Analysis of the resulting oxazolidine derivatives was performed using gas chromatography/mass spectrometry (GC/MS). The observed aldehyde derivatives included formaldehyde, acetaldehyde, propionaldehyde, acrolein, isobutyraldehyde, butyraldehyde, isovaleraldehyde, valeraldehyde, furfural, hexanal, cyclohexane carboxaldehyde and other unidentified compounds likely to be aldehyde isomers. Formaldehyde, acetaldehyde, and benzaldehyde were detected in blank samples. By comparison with an internal standard, a sample produced by drawing air with contaminants derived by heating rosin core solder through a sampling tube contained levels of formaldehyde and acetaldehyde much greater than seen in sampling tube blanks. Benzaldehyde was not shown to be present at a significantly greater level in samples from heating rosin core solder than in blanks prepared using the same analysis protocol. The use of National Institute of Occupational Safety and Health (NIOSH) method 2539 extraction procedures produced blanks with levels of formaldehyde significantly lower than with a modified extraction method (methylene chloride, no sonication). The modified extraction method produced significantly lower benzaldehyde levels in blanks compared with the NIOSH extraction method using toluene and sonication of sampling sorbent tubes.

Packed capillary column solvating gas chromatography of aldehydes

The rapid separation of aldehydes of environmental interest was investigated using packed capillary column solvating gas chromatography (SGC). Fused-silica capillary columns (250 μm I.D.) were packed with poly(dicyanoallylsiloxane)-encapsulated 10 μm spherical porous (80 A) silica particles. Excellent resolution and good peak shapes were achieved for underivatized aldehydes using carbon dioxide as mobile phase. The effects of temperature and pressure on column efficiency and selectivity were studied. Packed capillary SGC gave better resolution of ten standard aldehydes than open tubular gas chromatography using poly(biscyanopropylsiloxane) as stationary phase, and near-baseline resolution of these aldehydes was obtained in approximately 60 s.