Stephanie J. Wetzel

Post-source decay in the analysis of polystyrene by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

Various secondary series are observed in matrix-assisted laser desorption/ionization (MALDI) time-of-flight mass spectra of polystyrene. The number and positions of the series depend on the choice of matrix and added cation. For a given treatment, series observed in linear mode are not necessarily observed in reflectron mode, and vice versa. Post-source decay analysis was used to determine that the secondary series arise at least in part from formation and decay of adducts of polystyrene with matrix species. There is some treatment-to-treatment variation, but adduct formation and decay were observed for all tested treatments. The multiplicity of secondary series makes it unclear whether post-source decay occurs for the main series (polystyrene + cation)+ ions under the conditions normally used for polystyrene analysis. Such ions do undergo post-source decay at laser fluences greater than normally used. Although only polystyrene was investigated in this work, other polymers may also produce adduct and decay series in MALDI analysis. Their presence can mask the presence of minor components in a sample, but at least as observed here, do not have a strong influence on molecular mass determinations.

Development and validation of a hydrophilic interaction liquid chromatography-tandem mass spectrometry method for the quantification of lipid-related extracellular metabolites in Saccharomyces cerevisiae.

A highly sensitive hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) method was developed and validated for the quantification of glycerophosphoinositol (GroPIns), glycerophosphocholine (GroPCho), glycerol 3-phosphate (GroP), inositol, and choline in the extracellular medium of Saccharomyces cerevisiae. The media samples were pretreated with a single two-phase liquid extraction. Chromatographic separation was achieved on a Waters Xbridge HILIC (150 mm × 4.6 mm, 5 μm) column under isocratic conditions using a mobile phase composed of acetonitrile/water, 70:30 (v/v) with 10mM ammonium acetate (pH adjusted to 4.5) at a flow-rate of 0.5 mL/min. Using a triple quadrupole tandem mass spectrometer, samples were detected in multiple reaction monitoring (MRM) mode via an electrospray ionization (ESI) source. The calibration curves were linear (r² ≥ 0.995) over the range of 0.5-150 nM, with the lower limit of quantitation validated at 0.5 nM for all analytes. The intra- and inter-day precision (calculated by coefficient of variation, CV%) ranged from 1.24 to 5.88% and 2.46 to 9.77%, respectively, and intra- and inter-day accuracy (calculated by relative error, RE%) was between -8.42 to 8.22% and -9.35 to 6.62%, respectively, at all quality control levels. The extracellular metabolites were stable throughout various storage stability studies. The fully validated method was successfully applied to determine the extracellular levels of phospholipid-related metabolites in S. cerevisiae.

Improved sample preparation of glyphosate and methylphosphonic acid by EPA method 6800A and time-of-flight mass spectrometry using novel solid-phase extraction

The employment of chemical weapons by rogue states and/or terrorist organizations is an ongoing concern in the United States. The quantitative analysis of nerve agents must be rapid and reliable for use in the private and public sectors. Current methods describe a tedious and time-consuming derivatization for gas chromatography-mass spectrometry and liquid chromatography in tandem with mass spectrometry. Two solid-phase extraction (SPE) techniques for the analysis of glyphosate and methylphosphonic acid are described with the utilization of isotopically enriched analytes for quantitation via atmospheric pressure chemical ionization-quadrupole time-of-flight mass spectrometry (APCI-Q-TOF-MS) that does not require derivatization. Solid-phase extraction-isotope dilution mass spectrometry (SPE-IDMS) involves pre-equilibration of a naturally occurring sample with an isotopically enriched standard. The second extraction method, i-Spike, involves loading an isotopically enriched standard onto the SPE column before the naturally occurring sample. The sample and the spike are then co-eluted from the column enabling precise and accurate quantitation via IDMS. The SPE methods in conjunction with IDMS eliminate concerns of incomplete elution, matrix and sorbent effects, and MS drift. For accurate quantitation with IDMS, the isotopic contribution of all atoms in the target molecule must be statistically taken into account. This paper describes two newly developed sample preparation techniques for the analysis of nerve agent surrogates in drinking water as well as statistical probability analysis for proper molecular IDMS. The methods described in this paper demonstrate accurate molecular IDMS using APCI-Q-TOF-MS with limits of quantitation as low as 0.400 mg/kg for glyphosate and 0.031 mg/kg for methylphosphonic acid.

Sensitive and stable pre-calibrated solid-phase extraction columns for environmental and forensic quantification using isotope dilution mass spectrometry

A method was developed to pre-load solid-phase extraction columns (SPE) with isotopic calibrant for use with isotope dilution mass spectrometry (IDMS). The pre-calibration method was developed, optimized, and validated for the quantification of the pesticide glyphosate in drinking water using anion-exchange SPE, electrospray-ionization, and time-of-flight mass spectrometry (ESI-TOF-MS). The instrumental method obtained a mass-accuracy of 3 ppm and a limit of quantification (LOQ) of 0.3 ng mL−1. Quantification of glyphosate by IDMS significantly improved quantitative error and LOQ compared with the calibration curve. The pre-loading methodology was optimized for stability over time and validated in drinking water, exhibiting an accuracy of 1.25% ± 0.87% error with no significant difference from certified concentrations or traditional SPE. Method LOQ was 0.4 ng mL−1. Quantifying glyphosate in spiked drinking water sample produced high accuracy up to two-weeks after pre-loading columns, with an accuracy of 6.41% ± 7.10% error. A potential forensic application was investigated by adapting the pre-loading method to the quantification of seven drugs in synthetic urine using a mixed-mode SPE column. The ESI-TOF-MS method using traditional SPE produced accurate quantification of all seven drugs in synthetic urine with a mean error of 4.16% ± 3.07% and LOQ of 0.780 ng mL−1. The pre-loading method produced accurate quantification with 5.36% ± 4.73% error, with no significant difference from traditional SPE or certified standards. Five of the seven drugs were quantified at high accuracy one week after pre-loading, with 5.40% ± 4.57% error from certified values. This method may be applicable to analysts seeking to develop methods to improve the transfer of high-accuracy and precision methods between laboratories.